IP Library › Granted Patent US 12,612,605
Granted Patent B2
US 12,612,605 · App. 17/916,510 · Granted Apr 28, 2026

Carboxyesterase polypeptides for kinetic resolution

Inventors: Rama Voladri (Pleasanton, CA); Jessica Anna Hurtak (Mountain View, CA); Auric Anthony Sowell-Kantz (Mill Valley, CA); Nandhitha Subramanian (Cambridge, GB); Ericka Bermudez (Aptos, CA); David Entwistle (San Carlos, CA); Melissa Ann Mayo (Foster City, CA); James Nicholas Riggins (San Francisco, CA)
Assignee: Codexis, Inc.
C12N9/18
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,612,605
App. No.
17/916,510
Granted
Apr 28, 2026
Kind
B2
Abstract

The present invention provides engineered carboxyesterase enzymes having improved properties as compared to a naturally occurring wild-type carboxyesterase enzymes, as well as polynucleotides encoding the engineered carboxyesterase enzymes, host cells capable of expressing the engineered carboxyesterase enzymes, and methods of using the engineered carboxyesterase enzymes in hydrolysis reactions.

Claims (26)

1 . An engineered carboxyesterase comprising a polypeptide sequence having at least 90%, sequence identity to SEQ ID NO: 2 or a functional fragment thereof, wherein said engineered carboxyesterase comprises the substitution set 9/10/70 in said polypeptide sequence, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2, and wherein the engineered carboxyesterase comprises at least one improved property compared to the wild-type Xanthomonas campestris carboxyesterase of SEQ ID NO:2.

2 . The engineered carboxyesterase of claim 1 , wherein said engineered carboxyesterase further comprises one or more amino acid substitutions at positions selected from: X82, and X180.

3 . The engineered carboxyesterase of claim 1 , wherein said at least one substitution or substitution set in said polypeptide sequence further comprises substitutions at positions selected from: 3/4/6/11/36/38/82/180/200/214/229, 3/6/99, 3/8/11/13/82/107/229, 3/8/11/13/70/107, 3/8/11, 3/8/11/229, 3/8/13/82/107, 3/8/13/82/229, 3/8/82, 3/8/82/229, 3/8/229, 3/11/13/82, 3/11/782/107, 3/11/229, 3/82, 3/11, 3/9/11/82/180, 4/6/8/11/38/82/180/186/200/214/229, 4/6/11/36/82/180/200/214/229, 4/6/11/38/82/180/186/200/214, 4/6/82/229, 4/8/11/13/82/107/229, 4/8/11/36/38/82/180/200/214/229, 4/8/11/36/82/180/186/200/214/229, 4/8/11/82/180/200/214/229, 4/8/36/38/82/180/200/214/229, 4/11/36/38/82/180/200/214/229, 4/11/36/65/82/180/186/200/214, 4/11/36/82/180/200/214, 4/11/36/82/180/200/214/229, 4/11/82/180/200/214, 6/8/38/82/180/200/214, 6/11/36/82/180/200/214/229, 6/38/82/180/186/200/214/229, 6/82/180/186/200/214/229, 8, 8/11/13, 8/11/13/82, 8/11/13/82/107/229, 8/11/36/38/82/180/200/214, 8/11/38/82/180/200/214/229, 8/11/43/82/180/200/214, 8/11, 8/11/82/107/229, 8/11/229, 8/82/229, 11/12/49/51/73/82/180/200/225, 11/12/49/73/82/180, 11/12/51/54/73/82/108/148/180/225, 11/12/51/54/73/82/110/180, 11/12/51/54/73/82/148/180/225, 11/12/51/73/82/148/180, 11/12/51/73/82/148/180/200/225, 11/12/51/73/110/148/180, 11/17/49/82/180/200, 11/17/49/82/180/219, 11/17/51/54/82/148/180/200, 11/17/51/54/82/180, 11/17/73/82/87/180/219, 11/17/82/148/180/219, 11/35/82/180/200/214, 11/36/38/65/82/180/200/214/229, 11/36/38/82/180/200/214, 11/36/38/82/180/200/214/229, 11/36/82/180/196/214, 11/36/82/180/200/214, 11/36/82/180/200/214/229, 11/38/82/180/186/200/214, 11/40/78/82/180/200/214, 11/49/51/54/82/87/108/110/148/180/219, 11/49/51/54/82/87/108/180, 11/49/51/54/82/110/148/180, 11/49/51/82/108/10/148/180, 11/49/54/82/87/148/180, 11/49/82/87/108/110/180/200, 11/49/82/87/180, 11/49/82/110/148/180/225, 11/51/54/73/82/108/110/180, 11/51/54/73/82/108/180/200/225, 11/51/54/73/82/110/148/180, 11/51/54/82/148/180, 11/51/82/110/180/192/219, 11/54/82/87/108/180, 11/54/82/87/180/192, 11/54/82/148/180/200, 11/54/82/180/200/214, 11/70, 11/71/82/180/200/214, 9/10/11/73/82/87/110/180, 11/73/82/108/148/180, 11/73/82/148/180, 11/73/82/148/180/192/200, 11/73/82/180/200/214, 11/82, 11/82/87/110/180/219, 11/82/100/180/200/214, 11/82/107, 11/82/108/180/200/214, 11/82/148/180, 11/82/148/180/200/214, 11/82/180, 11/82/180/200/214, 11/87/180/200/214, 11/107, 12/49/51/54/82/108/148/180/225, 12/51/54/180/225, 12/51/73/82/148/180/200, 12/82/148/180/225, 12/82/180/200/214, 13, 13/82, 13/229, 13/207, 17, 17/49/51/54/73/82/108/110/148/180/192/200/219, 17/49/51/54/82/87/180/219, 17/49/82/148/180, 17/51/54/82/87/108/110/180/200, 17/51/73/82/87/108/110/148/180/219, 17/54/73/82/87/108/110/148/180/200, 17/54/73/82/108/110/148/180, 17/54/82/87/108/110/180, 17/73/82/87/180/219, 17/73/82/108/110/148/180/200, 17/82/87/148/180, 17/82/87/148/180/192/219, 17/82/108/148/180, 17/82/110/148/180, 17/82/110/148/180/200, 17/82/180/200/214, 17/104/160/165, 17/139/160/165, 26, 29, 36/82/180/200/214, 37, 38/82/180/200/214, 39, 39/186, 41, 45, 49/51/54/73/82/108/110/148/180/192, 49/51/54/73/82/148/180, 49/51/54/82/108/110/148/180/225, 49/51/54/70/82/110/148/180, 49/51/54/82/110/180, 49/51/54/82/148/180, 49/51/54/82/148/180/192, 49/51/82/108/148/180/219, 49/51/82/110/148/180, 49/51/82/180/225, 49/54/73/82/87/110/180, 49/54/82/87/110/180, 49/54/82/108/148/180/225, 49/73/82/87/108/148/180, 49/73/82/108/110/148/180/225, 49/82/87/148/180, 49/82/110/180/219, 51/54/73/82/108/180, 51/54/73/82/110/148/180, 51/54/73/82/110/180, 51/54/73/82/180/219, 51/54/73/82/180/225, 51/54/82/87/180, 51/54/82/148/180, 51/54/82/180, 51/54/82/180/200, 51/82/108/180, 51/82/110/180/200, 51/70/82/180/219, 54/73/82/108/148/180/192, 54/70/82/87/108/180/200, 54/70/82/180, 62, 73/82/87/108/110/180, 73/82/87/180/192/219, 73/82/87/180/200, 73/82/108/110/148/180, 73/82/108/110/180/200/219, 73/82/108/148/180, 73/82/108/148/180/192, 73/82/108/148/180/192/219, 73/82/108/180, 73/82/148/180, 73/82/148/180/200, 73/82/180/192, 73/82/180/192/219, 82, 82/87/108/110/148/180/192/219, 82/87/108/110/180, 82/87/108/110/180/192, 82/87/180, 82/107, 82/107/229, 82/108/110/180/219, 82/108/180, 82/110/148/180, 82/110/180/219, 82/148/180, 82/148/180/200, 82/148/180/200/219, 82/180, 82/180/192/219, 82/180/200/214, 82/229, 92, 99, 99/229, 107/229, 156, 165, 229, 8890, 96, 107, 110, 111, 112, 133, 178, 185, 186, 196, 211, 226, 11/24, 11/24/219, and 11/36/206/208, wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2.

4 . The engineered carboxyesterase of claim 1 , wherein the engineered carboxyesterase has at least 95% to SEQ ID NO: 2, and wherein said at least one substitution or substitution set in said polypeptide sequence comprises substitutions at positions selected from: 3/4/6/9/10/11/36/38/70/82/180/200/214/229, 3/6/9/10/70/99, 3/8/9/10/11/13/70/82/107/229, 3/8/9/10/11/13/70/107, 3/8/9/10/11/70, 3/8/9/10/11/70/229, 3/8/9/10/13/70/82/107, 3/8/9/10/13/70/82/229, 3/8/9/10/70/82, 3/8/9/10/70/82/229, 3/8/9/10/70/229, 3/9/10/11/13/82, 3/9/10/11/70/82/107, 3/9/10/11/70/229, 3/9/10/70/82, 3/9/11, 3/9/11/70/82/180, 4/6/8/9/10/11/38/70/82/180/186/200/214/229, 4/6/9/10/11/36/70/82/180/200/214/229, 4/6/9/10/11/38/70/82/180/186/200/214, 4/6/9/10/70/82/229, 4/8/9/10/11/13/70/82/107/229, 4/8/9/10/11/36/38/70/82/180/200/214/229, 4/8/9/10/11/36/70/82/180/186/200/214/229, 4/8/9/10/11/70/82/180/200/214/229, 4/8/9/10/36/38/70/82/180/200/214/229, 4/9/10/11/36/38/70/82/180/200/214/229, 4/9/10/11/36/65/70/82/180/186/200/214, 4/9/10/11/36/70/82/180/200/214, 4/9/10/11/36/70/82/180/200/214/229, 4/9/10/11/70/82/180/200/214, 6/8/9/10/38/70/82/180/200/214, 6/9/10/11/36/70/82/180/200/214/229, 6/9/10/38/70/82/180/186/200/214/229, 6/9/10/70/82/180/186/200/214/229, 8/9/10, 8/9/10/11/13/70, 8/9/10/11/13/70/82, 8/9/10/11/13/70/82/107/229, 8/9/10/11/36/38/70/82/180/200/214, 8/9/10/11/38/70/82/180/200/214/229, 8/9/10/11/43/70/82/180/200/214, 8/9/10/11/70, 8/9/10/11/70/82/107/229, 8/9/10/11/70/229, 8/9/10/70, 8/9/10/70/82/229, 9/10, 9/10/11/12/49/51/70/73/82/180/200/225, 9/10/11/12/49/70/73/82/180, 9/10/11/12/51/54/70/73/82/108/148/180/225, 9/10/11/12/51/54/70/73/82/110/180, 9/10/11/12/51/54/70/73/82/148/180/225, 9/10/11/12/51/70/73/82/148/180, 9/10/11/12/51/70/73/82/148/180/200/225, 9/10/11/12/51/70/73/110/148/180, 9/10/11/17/49/70/82/180/200, 9/10/11/17/49/70/82/180/219, 9/10/11/17/51/54/70/82/148/180/200, 9/10/11/17/51/54/70/82/180, 9/10/11/17/70/73/82/87/180/219, 9/10/11/17/70/82/148/180/219, 9/10/11/35/70/82/180/200/214, 9/10/11/36/38/65/70/82/180/200/214/229, 9/10/11/36/38/70/82/180/200/214, 9/10/11/36/38/70/82/180/200/214/229, 9/10/11/36/70/82/180/196/214, 9/10/11/36/70/82/180/200/214, 9/10/11/36/70/82/180/200/214/229, 9/10/11/38/70/82/180/186/200/214, 9/10/11/40/70/82/180/200/214, 9/10/11/49/51/54/70/82/87/108/110/148/180/219, 9/10/11/49/51/54/70/82/87/108/180, 9/10/11/49/51/54/70/82/110/148/180, 9/10/11/49/51/70/82/108/110/148/180, 9/10/11/49/54/70/82/87/148/180, 9/10/11/49/70/82/87/108/110/180/200, 9/10/11/49/70/82/87/180, 9/10/11/49/70/82/110/148/180/225, 9/10/11/51/54/70/73/82/108/110/180, 9/10/11/51/54/70/73/82/108/180/200/225, 9/10/11/51/54/70/73/82/110/148/180, 9/10/11/51/54/70/82/148/180, 9/10/11/51/70/82/110/180/192/219, 9/10/11/54/70/82/87/108/180, 9/10/11/54/70/82/87/180/192, 9/10/11/54/70/82/148/180/200, 9/10/11/54/70/82/180/200/214, 9/10/11/70, 9/10/11/70/71/82/180/200/214, 9/10/11/70/73/82/87/110/180, 9/10/11/70/73/82/108/148/180, 9/10/11/70/73/82/148/180, 9/10/11/70/73/82/148/180/192/200, 9/10/11/70/73/82/180/200/214, 9/10/11/70/82, 9/10/11/70/82/87/110/180/219, 9/10/11/70/82/100/180/200/214, 9/10/11/70/82/107, 9/10/11/70/82/108/180/200/214, 9/10/11/70/82/148/180, 9/10/11/70/82/148/180/200/214, 9/10/11/70/82/180, 9/10/11/70/82/180/200/214, 9/10/11/70/87/180/200/214, 9/10/11/70/107, 9/10/12/49/51/54/70/82/108/148/180/225, 9/10/12/51/54/70/180/225, 9/10/12/51/70/73/82/148/180/200, 9/10/12/70/82/148/180/225, 9/10/12/70/82/180/200/214, 9/10/13/70, 9/10/13/70/82, 9/10/13/70/229, 9/10/13/207, 9/10/17, 9/10/17/49/51/54/70/73/82/108/110/148/180/192/200/219, 9/10/17/49/51/54/70/82/87/180/219, 9/10/17/49/70/82/148/180, 9/10/17/51/54/70/82/87/108/110/180/200, 9/10/17/51/70/73/82/87/108/110/148/180/219, 9/10/17/54/70/73/82/87/108/110/148/180/200, 9/10/17/54/70/73/82/108/110/148/180, 9/10/17/54/70/82/87/108/110/180, 9/10/17/70/73/82/87/180/219, 9/10/17/70/73/82/108/110/148/180/200, 9/10/17/70/82/87/148/180, 9/10/17/70/82/87/148/180/192/219, 9/10/17/70/82/108/148/180, 9/10/17/70/82/110/148/180, 9/10/17/70/82/110/148/180/200, 9/10/17/70/82/180/200/214, 9/10/17/70/104/160/165, 9/10/17/70/139/160/165, 9/10/26, 9/10/29/70, 9/10/36/70/82/180/200/214, 9/10/37, 9/10/38/70/82/180/200/214, 9/10/39, 9/10/39/186, 9/10/41, 9/10/45/70, 9/10/49/51/54/70/73/82/108/110/148/180/192, 9/10/49/51/54/70/73/82/148/180, 9/10/49/51/54/70/82/108/110/148/180/225, 9/10/49/51/54/70/82/110/148/180, 9/10/49/51/54/70/82/110/180, 9/10/49/51/54/70/82/148/180, 9/10/49/51/54/70/82/148/180/192, 9/10/49/51/70/82/108/148/180/219, 9/10/49/51/70/82/110/148/180, 9/10/49/51/70/82/180/225, 9/10/49/54/70/73/82/87/110/180, 9/10/49/54/70/82/87/110/180, 9/10/49/54/70/82/108/148/180/225, 9/10/49/70/73/82/87/108/148/180, 9/10/49/70/73/82/108/110/148/180/225, 9/10/49/70/82/87/148/180, 9/10/49/70/82/110/180/219, 9/10/51/54/70/73/82/108/180, 9/10/51/54/70/73/82/110/148/180, 9/10/51/54/70/73/82/110/180, 9/10/51/54/70/73/82/180/219, 9/10/51/54/70/73/82/180/225, 9/10/51/54/70/82/87/180, 9/10/51/54/70/82/148/180, 9/10/51/54/70/82/180, 9/10/51/54/70/82/180/200, 9/10/51/70/82/108/180, 9/10/51/70/82/110/180/200, 9/10/51/70/82/180/219, 9/10/54/70/73/82/108/148/180/192, 9/10/54/70/82/87/108/180/200, 9/10/54/70/82/180, 9/10/62, 9/10/70, 9/10/70/73/82/87/108/110/180, 9/10/70/73/82/87/180/192/219, 9/10/70/73/82/87/180/200, 9/10/70/73/82/108/110/148/180, 9/10/70/73/82/108/110/180/200/219, 9/10/70/73/82/108/148/180, 9/10/70/73/82/108/148/180/192, 9/10/70/73/82/108/148/180/192/219, 9/10/70/73/82/108/180, 9/10/70/73/82/148/180, 9/10/70/73/82/148/180/200, 9/10/70/73/82/180/192, 9/10/70/73/82/180/192/219, 9/10/70/82, 9/10/70/82/87/108/110/148/180/192/219, 9/10/70/82/87/108/110/180, 9/10/70/82/87/108/110/180/192, 9/10/70/82/87/180, 9/10/70/82/107, 9/10/70/82/107/229, 9/10/70/82/108/110/180/219, 9/10/70/82/108/180, 9/10/70/82/110/148/180, 9/10/70/82/110/180/219, 9/10/70/82/148/180, 9/10/70/82/148/180/200, 9/10/70/82/148/180/200/219, 9/10/70/82/180, 9/10/70/82/180/192/219, 9/10/70/82/180/200/214, 9/10/70/82/229, 9/10/70/92, 9/10/70/99, 9/10/70/99/229, 9/10/70/107/229, 9/10/70/156, 9/10/70/165, 9/10/70/229, 9/10/88, 9/10/90, 9/10/96, 9/10/107, 9/10/110, 9/10/111, 9/10/112, 9/10/133, 9/10/178, 9/10/185, 9/10/186, 9/10/196, 9/10/211, 9/10/226, 11/24, 11/24/219, and 11/36/206/208, wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 82.

5 . The engineered carboxyesterase of claim 1 , wherein said at least one substitution or substitution set in said polypeptide sequence comprises further substitutions at positions selected from: 3/4/6/9/10/11/36/38/70/82/180/200/214/229, 3/6/9/10/70/99, 3/8/9/10/11/13/70/82/107/229, 3/8/9/10/11/13/70/107, 3/8/9/10/11/70, 3/8/9/10/11/70/229, 3/8/9/10/13/70/82/107, 3/8/9/10/13/70/82/229, 3/8/9/10/70/82, 3/8/9/10/70/82/229, 3/8/9/10/70/229, 3/9/10/11/13/82, 3/9/10/11/70/82/107, 3/9/10/11/70/229, 3/9/10/70/82, 3/9/11, 3/9/11/70/82/180, 4/6/8/9/10/11/38/70/82/180/186/200/214/229, 4/6/9/10/11/36/70/82/180/200/214/229, 4/6/9/10/11/38/70/82/180/186/200/214, 4/6/9/10/70/82/229, 4/8/9/10/11/13/70/82/107/229, 4/8/9/10/11/36/38/70/82/180/200/214/229, 4/8/9/10/11/36/70/82/180/186/200/214/229, 4/8/9/10/11/70/82/180/200/214/229, 4/8/9/10/36/38/70/82/180/200/214/229, 4/9/10/11/36/38/70/82/180/200/214/229, 4/9/10/11/36/65/70/82/180/186/200/214, 4/9/10/11/36/70/82/180/200/214, 4/9/10/11/36/70/82/180/200/214/229, 4/9/10/11/70/82/180/200/214, 6/8/9/10/38/70/82/180/200/214, 6/9/10/11/36/70/82/180/200/214/229, 6/9/10/38/70/82/180/186/200/214/229, 6/9/10/70/82/180/186/200/214/229, 8/9/10, 8/9/10/11/13/70, 8/9/10/11/13/70/82, 8/9/10/11/13/70/82/107/229, 8/9/10/11/36/38/70/82/180/200/214, 8/9/10/11/38/70/82/180/200/214/229, 8/9/10/11/43/70/82/180/200/214, 8/9/10/11/70, 8/9/10/11/70/82/107/229, 8/9/10/11/70/229, 8/9/10/70, 8/9/10/70/82/229, 9/10, 9/10/11/12/49/51/70/73/82/180/200/225, 9/10/11/12/49/70/73/82/180, 9/10/11/12/51/54/70/73/82/108/148/180/225, 9/10/11/12/51/54/70/73/82/110/180, 9/10/11/12/51/54/70/73/82/148/180/225, 9/10/11/12/51/70/73/82/148/180, 9/10/11/12/51/70/73/82/148/180/200/225, 9/10/11/12/51/70/73/110/148/180, 9/10/11/17/49/70/82/180/200, 9/10/11/17/49/70/82/180/219, 9/10/11/17/51/54/70/82/148/180/200, 9/10/11/17/51/54/70/82/180, 9/10/11/17/70/73/82/87/180/219, 9/10/11/17/70/82/148/180/219, 9/10/11/35/70/82/180/200/214, 9/10/11/36/38/65/70/82/180/200/214/229, 9/10/11/36/38/70/82/180/200/214, 9/10/11/36/38/70/82/180/200/214/229, 9/10/11/36/70/82/180/196/214, 9/10/11/36/70/82/180/200/214, 9/10/11/36/70/82/180/200/214/229, 9/10/11/38/70/82/180/186/200/214, 9/10/11/40/70/82/180/200/214, 9/10/11/49/51/54/70/82/87/108/110/148/180/219, 9/10/11/49/51/54/70/82/87/108/180, 9/10/11/49/51/54/70/82/110/148/180, 9/10/11/49/51/70/82/108/110/148/180, 9/10/11/49/54/70/82/87/148/180, 9/10/11/49/70/82/87/108/110/180/200, 9/10/11/49/70/82/87/180, 9/10/11/49/70/82/110/148/180/225, 9/10/11/51/54/70/73/82/108/110/180, 9/10/11/51/54/70/73/82/108/180/200/225, 9/10/11/51/54/70/73/82/110/148/180, 9/10/11/51/54/70/82/148/180, 9/10/11/51/70/82/110/180/192/219, 9/10/11/54/70/82/87/108/180, 9/10/11/54/70/82/87/180/192, 9/10/11/54/70/82/148/180/200, 9/10/11/54/70/82/180/200/214, 9/10/11/70, 9/10/11/70/71/82/180/200/214, 9/10/11/70/73/82/87/110/180, 9/10/11/70/73/82/108/148/180, 9/10/11/70/73/82/148/180, 9/10/11/70/73/82/148/180/192/200, 9/10/11/70/73/82/180/200/214, 9/10/11/70/82, 9/10/11/70/82/87/110/180/219, 9/10/11/70/82/100/180/200/214, 9/10/11/70/82/107, 9/10/11/70/82/108/180/200/214, 9/10/11/70/82/148/180, 9/10/11/70/82/148/180/200/214, 9/10/11/70/82/180, 9/10/11/70/82/180/200/214, 9/10/11/70/87/180/200/214, 9/10/11/70/107, 9/10/12/49/51/54/70/82/108/148/180/225, 9/10/12/51/54/70/180/225, 9/10/12/51/70/73/82/148/180/200, 9/10/12/70/82/148/180/225, 9/10/12/70/82/180/200/214, 9/10/13/70, 9/10/13/70/82, 9/10/13/70/229, 9/10/13/207, 9/10/17, 9/10/17/49/51/54/70/73/82/108/110/148/180/192/200/219, 9/10/17/49/51/54/70/82/87/180/219, 9/10/17/49/70/82/148/180, 9/10/17/51/54/70/82/87/108/110/180/200, 9/10/17/51/70/73/82/87/108/110/148/180/219, 9/10/17/54/70/73/82/87/108/110/148/180/200, 9/10/17/54/70/73/82/108/110/148/180, 9/10/17/54/70/82/87/108/110/180, 9/10/17/70/73/82/87/180/219, 9/10/17/70/73/82/108/110/148/180/200, 9/10/17/70/82/87/148/180, 9/10/17/70/82/87/148/180/192/219, 9/10/17/70/82/108/148/180, 9/10/17/70/82/110/148/180, 9/10/17/70/82/110/148/180/200, 9/10/17/70/82/180/200/214, 9/10/17/70/104/160/165, 9/10/17/70/139/160/165, 9/10/26, 9/10/29/70, 9/10/36/70/82/180/200/214, 9/10/37, 9/10/38/70/82/180/200/214, 9/10/39, 9/10/39/186, 9/10/41, 9/10/45/70, 9/10/49/51/54/70/73/82/108/110/148/180/192, 9/10/49/51/54/70/73/82/148/180, 9/10/49/51/54/70/82/108/110/148/180/225, 9/10/49/51/54/70/82/110/148/180, 9/10/49/51/54/70/82/110/180, 9/10/49/51/54/70/82/148/180, 9/10/49/51/54/70/82/148/180/192, 9/10/49/51/70/82/108/148/180/219, 9/10/49/51/70/82/110/148/180, 9/10/49/51/70/82/180/225, 9/10/49/54/70/73/82/87/110/180, 9/10/49/54/70/82/87/110/180, 9/10/49/54/70/82/108/148/180/225, 9/10/49/70/73/82/87/108/148/180, 9/10/49/70/73/82/108/110/148/180/225, 9/10/49/70/82/87/148/180, 9/10/49/70/82/110/180/219, 9/10/51/54/70/73/82/108/180, 9/10/51/54/70/73/82/110/148/180, 9/10/51/54/70/73/82/110/180, 9/10/51/54/70/73/82/180/219, 9/10/51/54/70/73/82/180/225, 9/10/51/54/70/82/87/180, 9/10/51/54/70/82/148/180, 9/10/51/54/70/82/180, 9/10/51/54/70/82/180/200, 9/10/51/70/82/108/180, 9/10/51/70/82/110/180/200, 9/10/51/70/82/180/219, 9/10/54/70/73/82/108/148/180/192, 9/10/54/70/82/87/108/180/200, 9/10/54/70/82/180, 9/10/62, 9/10/70, 9/10/70/73/82/87/108/110/180, 9/10/70/73/82/87/180/192/219, 9/10/70/73/82/87/180/200, 9/10/70/73/82/108/110/148/180, 9/10/70/73/82/108/110/180/200/219, 9/10/70/73/82/108/148/180, 9/10/70/73/82/108/148/180/192, 9/10/70/73/82/108/148/180/192/219, 9/10/70/73/82/108/180, 9/10/70/73/82/148/180, 9/10/70/73/82/148/180/200, 9/10/70/73/82/180/192, 9/10/70/73/82/180/192/219, 9/10/70/82, 9/10/70/82/87/108/110/148/180/192/219, 9/10/70/82/87/108/110/180, 9/10/70/82/87/108/110/180/192, 9/10/70/82/87/180, 9/10/70/82/107, 9/10/70/82/107/229, 9/10/70/82/108/110/180/219, 9/10/70/82/108/180, 9/10/70/82/110/148/180, 9/10/70/82/110/180/219, 9/10/70/82/148/180, 9/10/70/82/148/180/200, 9/10/70/82/148/180/200/219, 9/10/70/82/180, 9/10/70/82/180/192/219, 9/10/70/82/180/200/214, 9/10/70/82/229, 9/10/70/92, 9/10/70/99, 9/10/70/99/229, 9/10/70/107/229, 9/10/70/156, 9/10/70/165, 9/10/70/229, 9/10/88, 9/10/90, 9/10/96, 9/10/107, 9/10/110, 9/10/111, 9/10/112, 9/10/133, 9/10/178, 9/10/185, 9/10/186, 9/10/196, 9/10/211, 9/10/226, 11/24, 11/24/219, and 11/36/206/208, wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 268.

6 . The engineered carboxyesterase of claim 1 , wherein said at least one substitution or substitution set in said polypeptide sequence further comprises substitutions at positions selected from: 3/4/6/9/10/11/36/38/70/82/180/200/214/229, 3/6/9/10/70/99, 3/8/9/10/11/13/70/82/107/229, 3/8/9/10/11/13/70/107, 3/8/9/10/11/70, 3/8/9/10/11/70/229, 3/8/9/10/13/70/82/107, 3/8/9/10/13/70/82/229, 3/8/9/10/70/82, 3/8/9/10/70/82/229, 3/8/9/10/70/229, 3/9/10/11/13/82, 3/9/10/11/70/82/107, 3/9/10/11/70/229, 3/9/10/70/82, 3/9/11, 3/9/11/70/82/180, 4/6/8/9/10/11/38/70/82/180/186/200/214/229, 4/6/9/10/11/36/70/82/180/200/214/229, 4/6/9/10/11/38/70/82/180/186/200/214, 4/6/9/10/70/82/229, 4/8/9/10/11/13/70/82/107/229, 4/8/9/10/11/36/38/70/82/180/200/214/229, 4/8/9/10/11/36/70/82/180/186/200/214/229, 4/8/9/10/11/70/82/180/200/214/229, 4/8/9/10/36/38/70/82/180/200/214/229, 4/9/10/11/36/38/70/82/180/200/214/229, 4/9/10/11/36/65/70/82/180/186/200/214, 4/9/10/11/36/70/82/180/200/214, 4/9/10/11/36/70/82/180/200/214/229, 4/9/10/11/70/82/180/200/214, 6/8/9/10/38/70/82/180/200/214, 6/9/10/11/36/70/82/180/200/214/229, 6/9/10/38/70/82/180/186/200/214/229, 6/9/10/70/82/180/186/200/214/229, 8/9/10, 8/9/10/11/13/70, 8/9/10/11/13/70/82, 8/9/10/11/13/70/82/107/229, 8/9/10/11/36/38/70/82/180/200/214, 8/9/10/11/38/70/82/180/200/214/229, 8/9/10/11/43/70/82/180/200/214, 8/9/10/11/70, 8/9/10/11/70/82/107/229, 8/9/10/11/70/229, 8/9/10/70, 8/9/10/70/82/229, 9/10, 9/10/11/12/49/51/70/73/82/180/200/225, 9/10/11/12/49/70/73/82/180, 9/10/11/12/51/54/70/73/82/108/148/180/225, 9/10/11/12/51/54/70/73/82/110/180, 9/10/11/12/51/54/70/73/82/148/180/225, 9/10/11/12/51/70/73/82/148/180, 9/10/11/12/51/70/73/82/148/180/200/225, 9/10/11/12/51/70/73/110/148/180, 9/10/11/17/49/70/82/180/200, 9/10/11/17/49/70/82/180/219, 9/10/11/17/51/54/70/82/148/180/200, 9/10/11/17/51/54/70/82/180, 9/10/11/17/70/73/82/87/180/219, 9/10/11/17/70/82/148/180/219, 9/10/11/35/70/82/180/200/214, 9/10/11/36/38/65/70/82/180/200/214/229, 9/10/11/36/38/70/82/180/200/214, 9/10/11/36/38/70/82/180/200/214/229, 9/10/11/36/70/82/180/196/214, 9/10/11/36/70/82/180/200/214, 9/10/11/36/70/82/180/200/214/229, 9/10/11/38/70/82/180/186/200/214, 9/10/11/40/70/82/180/200/214, 9/10/11/49/51/54/70/82/87/108/110/148/180/219, 9/10/11/49/51/54/70/82/87/108/180, 9/10/11/49/51/54/70/82/110/148/180, 9/10/11/49/51/70/82/108/110/148/180, 9/10/11/49/54/70/82/87/148/180, 9/10/11/49/70/82/87/108/110/180/200, 9/10/11/49/70/82/87/180, 9/10/11/49/70/82/110/148/180/225, 9/10/11/51/54/70/73/82/108/110/180, 9/10/11/51/54/70/73/82/108/180/200/225, 9/10/11/51/54/70/73/82/110/148/180, 9/10/11/51/54/70/82/148/180, 9/10/11/51/70/82/110/180/192/219, 9/10/11/54/70/82/87/108/180, 9/10/11/54/70/82/87/180/192, 9/10/11/54/70/82/148/180/200, 9/10/11/54/70/82/180/200/214, 9/10/11/70, 9/10/11/70/71/82/180/200/214, 9/10/11/70/73/82/87/110/180, 9/10/11/70/73/82/108/148/180, 9/10/11/70/73/82/148/180, 9/10/11/70/73/82/148/180/192/200, 9/10/11/70/73/82/180/200/214, 9/10/11/70/82, 9/10/11/70/82/87/110/180/219, 9/10/11/70/82/100/180/200/214, 9/10/11/70/82/107, 9/10/11/70/82/108/180/200/214, 9/10/11/70/82/148/180, 9/10/11/70/82/148/180/200/214, 9/10/11/70/82/180, 9/10/11/70/82/180/200/214, 9/10/11/70/87/180/200/214, 9/10/11/70/107, 9/10/12/49/51/54/70/82/108/148/180/225, 9/10/12/51/54/70/180/225, 9/10/12/51/70/73/82/148/180/200, 9/10/12/70/82/148/180/225, 9/10/12/70/82/180/200/214, 9/10/13/70, 9/10/13/70/82, 9/10/13/70/229, 9/10/13/207, 9/10/17, 9/10/17/49/51/54/70/73/82/108/110/148/180/192/200/219, 9/10/17/49/51/54/70/82/87/180/219, 9/10/17/49/70/82/148/180, 9/10/17/51/54/70/82/87/108/110/180/200, 9/10/17/51/70/73/82/87/108/110/148/180/219, 9/10/17/54/70/73/82/87/108/110/148/180/200, 9/10/17/54/70/73/82/108/110/148/180, 9/10/17/54/70/82/87/108/110/180, 9/10/17/70/73/82/87/180/219, 9/10/17/70/73/82/108/110/148/180/200, 9/10/17/70/82/87/148/180, 9/10/17/70/82/87/148/180/192/219, 9/10/17/70/82/108/148/180, 9/10/17/70/82/110/148/180, 9/10/17/70/82/110/148/180/200, 9/10/17/70/82/180/200/214, 9/10/17/70/104/160/165, 9/10/17/70/139/160/165, 9/10/26, 9/10/29/70, 9/10/36/70/82/180/200/214, 9/10/37, 9/10/38/70/82/180/200/214, 9/10/39, 9/10/39/186, 9/10/41, 9/10/45/70, 9/10/49/51/54/70/73/82/108/110/148/180/192, 9/10/49/51/54/70/73/82/148/180, 9/10/49/51/54/70/82/108/110/148/180/225, 9/10/49/51/54/70/82/110/148/180, 9/10/49/51/54/70/82/110/180, 9/10/49/51/54/70/82/148/180, 9/10/49/51/54/70/82/148/180/192, 9/10/49/51/70/82/108/148/180/219, 9/10/49/51/70/82/110/148/180, 9/10/49/51/70/82/180/225, 9/10/49/54/70/73/82/87/110/180, 9/10/49/54/70/82/87/110/180, 9/10/49/54/70/82/108/148/180/225, 9/10/49/70/73/82/87/108/148/180, 9/10/49/70/73/82/108/110/148/180/225, 9/10/49/70/82/87/148/180, 9/10/49/70/82/110/180/219, 9/10/51/54/70/73/82/108/180, 9/10/51/54/70/73/82/110/148/180, 9/10/51/54/70/73/82/110/180, 9/10/51/54/70/73/82/180/219, 9/10/51/54/70/73/82/180/225, 9/10/51/54/70/82/87/180, 9/10/51/54/70/82/148/180, 9/10/51/54/70/82/180, 9/10/51/54/70/82/180/200, 9/10/51/70/82/108/180, 9/10/51/70/82/110/180/200, 9/10/51/70/82/180/219, 9/10/54/70/73/82/108/148/180/192, 9/10/54/70/82/87/108/180/200, 9/10/54/70/82/180, 9/10/62, 9/10/70, 9/10/70/73/82/87/108/110/180, 9/10/70/73/82/87/180/192/219, 9/10/70/73/82/87/180/200, 9/10/70/73/82/108/110/148/180, 9/10/70/73/82/108/110/180/200/219, 9/10/70/73/82/108/148/180, 9/10/70/73/82/108/148/180/192, 9/10/70/73/82/108/148/180/192/219, 9/10/70/73/82/108/180, 9/10/70/73/82/148/180, 9/10/70/73/82/148/180/200, 9/10/70/73/82/180/192, 9/10/70/73/82/180/192/219, 9/10/70/82, 9/10/70/82/87/108/110/148/180/192/219, 9/10/70/82/87/108/110/180, 9/10/70/82/87/108/110/180/192, 9/10/70/82/87/180, 9/10/70/82/107, 9/10/70/82/107/229, 9/10/70/82/108/110/180/219, 9/10/70/82/108/180, 9/10/70/82/110/148/180, 9/10/70/82/110/180/219, 9/10/70/82/148/180, 9/10/70/82/148/180/200, 9/10/70/82/148/180/200/219, 9/10/70/82/180, 9/10/70/82/180/192/219, 9/10/70/82/180/200/214, 9/10/70/82/229, 9/10/70/92, 9/10/70/99, 9/10/70/99/229, 9/10/70/107/229, 9/10/70/156, 9/10/70/165, 9/10/70/229, 9/10/88, 9/10/90, 9/10/96, 9/10/107, 9/10/110, 9/10/111, 9/10/112, 9/10/133, 9/10/178, 9/10/185, 9/10/186, 9/10/196, 9/10/211, 9/10/226, 11/24, 11/24/219, and 11/36/206/208, wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 536.

7 . The engineered carboxyesterase of claim 1 , wherein said polypeptide sequence is selected from the even-numbered sequence identifiers of: SEQ ID NOs: 6-160, 164-194, 198-228, 232-240, 246-262, 266-274, 278, 286-298, 300-310, 314-322, 326, 328, 332-348, 352, 356-374, 378-392, 398-408, 412, 416-444, 450, 452, 456-462, 466-470, 474-480, 484-490, 494, 496, 502-506, 510-516, 520, 524, 528-532 and 536.

8 . The engineered carboxyesterase of claim 1 , wherein said improved property is selected from: improved hydrolysis activity, solvent tolerance, thermostability, pH stability, enantioselectivity, regioselectivity, stereoselectivity, substrate scope, and/or reduced substrate or product inhibition, and reduced toxicity to bacterial host cells producing said engineered carboxyesterase.

9 . The engineered carboxyesterase of claim 8 , wherein said bacterial host cells comprise E. coli.

10 . The engineered carboxyesterase of claim 1 , wherein said engineered carboxyesterase exhibits greater activity than wild-type Xanthomonas campestris carboxyesterase in the conversion of compound (2a)

to compound (3a)

11 . The engineered carboxyesterase of claim 1 , wherein said engineered carboxyesterase exhibits greater enantioselectivity than wild-type Xanthomonas campestris carboxyesterase in the conversion of compound (2a)

to compound (3a)

12 . The engineered carboxyesterase of claim 1 , wherein said engineered carboxyesterase is purified.

13 . The engineered carboxyesterase of claim 1 , wherein said engineered carboxyesterase is immobilized.

14 . A composition comprising at least one engineered carboxyesterase of claim 1 .

15 . A polynucleotide sequence encoding at least one engineered carboxyesterase of claim 1 .

16 . The polynucleotide sequence of claim 15 , wherein said polynucleotide sequence is operably linked to a control sequence.

17 . The polynucleotide sequence of claim 15 , wherein said polynucleotide sequence is codon optimized.

18 . An expression vector comprising at least one polynucleotide sequence of claim 15 .

19 . A host cell comprising at least one expression vector of claim 18 .

20 . A host cell comprising at least one polynucleotide sequence of claim 15 .

21 . The host cell of claim 19 , wherein said host cell is E. coli.

22 . A method of producing an engineered carboxyesterase in a host cell, comprising culturing the host cell of claim 19 , under suitable conditions, such that at least one engineered carboxyesterase is produced.

23 . The method of claim 22 , further comprising recovering at least one engineered carboxyesterase from the culture and/or host cell.

24 . The method of claim 23 , further comprising the step of purifying said at least one engineered carboxyesterase.

Assignments (3)
SECURITY INTEREST Recorded Feb 15, 2024
From: CODEXIS, INC.
To: INNOVATUS LIFE SCIENCES LENDING FUND I, LP, AS COLLATERAL AGENT
Reel/Frame 066600/0650 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 9, 2024
From: VOLADRI, RAMA; HURTAK, JESSICA ANNA; SUBRAMANIAN, NANDHITHA; BERMUDEZ, ERICKA; RIGGINS, JAMES NICHOLAS; SOWELL-KANTZ, AURIC ANTHONY; ENTWISTLE, DAVID; MAYO, MELISSA; NOVICK, SCOTT J.
To: CODEXIS, INC.
Reel/Frame 066432/0066 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 8, 2024
From: VOLADRI, RAMA; HURTAK, JESSICA ANNA; SOWELL-KANTZ, AURIC ANTHONY; ENTWISTLE, DAVID; MAYO, MELISSA ANN; RIGGINS, JAMES NICHOLAS; NOVICK, SCOTT J.; BERMUDEZ, ERICKA; SUBRAMANIAN, NANDHITHA
To: CODEXIS, INC.
Reel/Frame 066414/0572 →
Continuity (2)
Provisional Application 63008061 · Apr 10, 2020
Related Publication 20230159904A1 · May 25, 2023
References Cited (198)
US 5605793A · Stemmer · 1997 [cited by applicant]
US 5811238A · Stemmer et al. · 1998 [cited by applicant]
US 5830721A · Stemmer et al. · 1998 [cited by applicant]
US 5834252A · Stemmer et al. · 1998 [cited by applicant]
US 5837458A · Minshull et al. · 1998 [cited by applicant]
US 5928905A · Stemmer et al. · 1999 [cited by applicant]
US 6096548A · Stemmer · 2000 [cited by applicant]
US 6117679A · Stemmer · 2000 [cited by applicant]
US 6132970A · Stemmer · 2000 [cited by applicant]
US 6165793A · Stemmer · 2000 [cited by applicant]
US 6180406B1 · Stemmer · 2001 [cited by applicant]
US 6251674B1 · Tobin et al. · 2001 [cited by applicant]
US 6265201B1 · Wackett et al. · 2001 [cited by applicant]
US 6277638B1 · Stemmer · 2001 [cited by applicant]
US 6287861B1 · Stemmer et al. · 2001 [cited by applicant]
US 6287862B1 · delCardayre et al. · 2001 [cited by applicant]
US 6291242B1 · Stemmer · 2001 [cited by applicant]
US 6297053B1 · Stemmer · 2001 [cited by applicant]
US 6303344B1 · Patten et al. · 2001 [cited by applicant]
US 6309883B1 · Minshull et al. · 2001 [cited by applicant]
US 6319713B1 · Patten et al. · 2001 [cited by applicant]
US 6319714B1 · Crameri et al. · 2001 [cited by applicant]
US 6323030B1 · Stemmer · 2001 [cited by applicant]
US 6326204B1 · delCardayre et al. · 2001 [cited by applicant]
US 6335160B1 · Patten et al. · 2002 [cited by applicant]
US 6335198B1 · delCardayre et al. · 2002 [cited by applicant]
US 6337186B1 · Krebber · 2002 [cited by applicant]
US 6344356B1 · Stemmer · 2002 [cited by applicant]
US 6352859B1 · delCardayre et al. · 2002 [cited by applicant]
US 6355484B1 · Patten et al. · 2002 [cited by applicant]
US 6358740B1 · Patten et al. · 2002 [cited by applicant]
US 6358742B1 · Stemmer · 2002 [cited by applicant]
US 6365377B1 · Patten et al. · 2002 [cited by applicant]
US 6365408B1 · Stemmer · 2002 [cited by applicant]
US 6368861B1 · Crameri et al. · 2002 [cited by applicant]
US 6372497B1 · Stemmer · 2002 [cited by applicant]
US 6376246B1 · Crameri et al. · 2002 [cited by applicant]
US 6379964B1 · delCardayre et al. · 2002 [cited by applicant]
US 6387702B1 · Stemmer · 2002 [cited by applicant]
US 6391552B2 · Stemmer · 2002 [cited by applicant]
US 6391640B1 · Minshull et al. · 2002 [cited by applicant]
US 6395547B1 · Stemmer · 2002 [cited by applicant]
US 6406855B1 · Patten et al. · 2002 [cited by applicant]
US 6406910B1 · Patten et al. · 2002 [cited by applicant]
US 6413745B1 · Patten et al. · 2002 [cited by applicant]
US 6413774B1 · Stemmer · 2002 [cited by applicant]
US 6420175B1 · Stemmer · 2002 [cited by applicant]
US 6423542B1 · Crameri et al. · 2002 [cited by applicant]
US 6426224B1 · Crameri et al. · 2002 [cited by applicant]
US 6436675B1 · Welch et al. · 2002 [cited by applicant]
US 6444468B1 · Stemmer et al. · 2002 [cited by applicant]
US 6455253B1 · Patten et al. · 2002 [cited by applicant]
US 6479652B1 · Crameri et al. · 2002 [cited by applicant]
US 6482647B1 · Stemmer · 2002 [cited by applicant]
US 6483011B1 · Stemmer et al. · 2002 [cited by applicant]
US 6484105B2 · Zhang · 2002 [cited by applicant]
US 6489146B2 · Stemmer et al. · 2002 [cited by applicant]
US 6500617B1 · Stemmer et al. · 2002 [cited by applicant]
US 6500639B2 · Subramanian · 2002 [cited by applicant]
US 6506602B1 · Stemmer · 2003 [cited by applicant]
US 6506603B1 · Stemmer · 2003 [cited by applicant]
US 6518065B1 · Stemmer · 2003 [cited by applicant]
US 6519065B1 · Colbourne et al. · 2003 [cited by applicant]
US 6521453B1 · Crameri et al. · 2003 [cited by applicant]
US 6528311B1 · delCardayre et al. · 2003 [cited by applicant]
US 6537746B2 · Arnold et al. · 2003 [cited by applicant]
US 6573098B1 · Stemmer · 2003 [cited by applicant]
US 6576467B1 · Stemmer · 2003 [cited by applicant]
US 6579678B1 · Patten et al. · 2003 [cited by applicant]
US 6586182B1 · Patten et al. · 2003 [cited by applicant]
US 6602986B1 · Stemmer et al. · 2003 [cited by applicant]
US 6605430B1 · Affholter et al. · 2003 [cited by applicant]
US 6613514B2 · Patten et al. · 2003 [cited by applicant]
US 6653072B1 · Patten et al. · 2003 [cited by applicant]
US 6686515B1 · Lassner et al. · 2004 [cited by applicant]
US 6703240B1 · Stemmer et al. · 2004 [cited by applicant]
US 6716631B1 · delCardayre et al. · 2004 [cited by applicant]
US 6825001B2 · Wackett et al. · 2004 [cited by applicant]
US 6902922B2 · Ness et al. · 2005 [cited by applicant]
US 6917882B2 · Selifonov et al. · 2005 [cited by applicant]
US 6946296B2 · Patten et al. · 2005 [cited by applicant]
US 6961664B2 · Selifonov et al. · 2005 [cited by applicant]
US 6995017B1 · Stemmer · 2006 [cited by applicant]
US 7024312B1 · Selifonov et al. · 2006 [cited by applicant]
US 7058515B1 · Selifonov et al. · 2006 [cited by applicant]
US 7105297B2 · Minshull et al. · 2006 [cited by applicant]
US 7148054B2 · delCardayre et al. · 2006 [cited by applicant]
US 7220566B2 · Ness et al. · 2007 [cited by applicant]
US 7288375B2 · Stemmer et al. · 2007 [cited by applicant]
US 7384387B1 · Raillard et al. · 2008 [cited by applicant]
US 7421347B2 · Selifonov et al. · 2008 [cited by applicant]
US 7430477B2 · Selifonov et al. · 2008 [cited by applicant]
US 7462469B2 · Bass et al. · 2008 [cited by applicant]
US 7534564B2 · Patten et al. · 2009 [cited by applicant]
US 7620500B2 · Mundorff et al. · 2009 [cited by applicant]
US 7620502B2 · Selifonov et al. · 2009 [cited by applicant]
US 7629170B2 · delCardayre et al. · 2009 [cited by applicant]
US 7702464B1 · Emig et al. · 2010 [cited by applicant]
US 7747391B2 · Gustafsson et al. · 2010 [cited by applicant]
US 7747393B2 · Fox · 2010 [cited by applicant]
US 7751986B2 · Gustafsson et al. · 2010 [cited by applicant]
US 7776598B2 · Patten et al. · 2010 [cited by applicant]
US 7783428B2 · Gustafsson et al. · 2010 [cited by applicant]
US 7795030B2 · Minshull et al. · 2010 [cited by applicant]
US 7853410B2 · Selifonov et al. · 2010 [cited by applicant]
US 7868138B2 · Stemmer et al. · 2011 [cited by applicant]
US 7873477B1 · Gustafsson et al. · 2011 [cited by applicant]
US 7873499B2 · Selifonov et al. · 2011 [cited by applicant]
US 7904249B2 · Selifonov et al. · 2011 [cited by applicant]
US 7957912B2 · Selifonov et al. · 2011 [cited by applicant]
US 7981614B2 · Stemmer et al. · 2011 [cited by applicant]
US 8014961B2 · Bass et al. · 2011 [cited by applicant]
US 8029988B2 · Crameri et al. · 2011 [cited by applicant]
US 8048674B2 · Minshull et al. · 2011 [cited by applicant]
US 8058001B2 · Crameri et al. · 2011 [cited by applicant]
US 8076138B2 · delCardayre et al. · 2011 [cited by applicant]
US 8108150B2 · Mundorff et al. · 2012 [cited by applicant]
US 8170806B2 · Selifonov et al. · 2012 [cited by applicant]
US 8224580B2 · Mundorff et al. · 2012 [cited by applicant]
US 8377681B2 · delCardayre et al. · 2013 [cited by applicant]
US 8383346B2 · Colbeck et al. · 2013 [cited by applicant]
US 8457903B1 · Emig et al. · 2013 [cited by applicant]
US 8504498B2 · Fox · 2013 [cited by applicant]
US 8589085B2 · Selifonov et al. · 2013 [cited by applicant]
US 8762066B2 · Fox · 2014 [cited by applicant]
US 8768871B2 · Fox · 2014 [cited by applicant]
US 9453018B2 · Dilger et al. · 2016 [cited by applicant]
US 9593326B2 · Clark et al. · 2017 [cited by applicant]
US 9665694B2 · Cope · 2017 [cited by applicant]
US 9684771B2 · Cope et al. · 2017 [cited by applicant]
US 9714437B2 · Chan et al. · 2017 [cited by applicant]
US 20030233675A1 · Cao et al. · 2003 [cited by applicant]
US 20080220990A1 · Fox · 2008 [cited by applicant]
US 20080229451A1 · Cao et al. · 2008 [cited by applicant]
US 20090312196A1 · Colbeck et al. · 2009 [cited by applicant]
US 20160096839A1 · Dilger et al. · 2016 [cited by applicant]
US 20190177707A1 · Voladri et al. · 2019 [cited by applicant]
WO 9522625A1 · 1995 [cited by applicant]
WO 9533836A1 · 1995 [cited by applicant]
WO 9600787A1 · 1996 [cited by applicant]
WO 970078A1 · 1997 [cited by applicant]
WO 9735966A1 · 1997 [cited by applicant]
WO 9827230A1 · 1998 [cited by applicant]
WO 0042651A1 · 2000 [cited by applicant]
WO 0175767A2 · 2001 [cited by applicant]
WO 2007128496A2 · 2007 [cited by applicant]
WO 2009008908A2 · 2009 [cited by applicant]
WO 2009152336A1 · 2009 [cited by applicant]
WO 2019116311A1 · 2019 [cited by applicant]
WO 2019118557 · 2019 [cited by applicant]
B0RNI6_XANCB. UniProtKB Database. Jan. 16, 2019. [cited by examiner]
Fransceus. J Ind Microbiol Biotechnol. May 2017;44(4-5):687-695. [cited by examiner]
Sanavia. Computational and Structural Biotechnology Journal, vol. 18, 2020, pp. 1968-1979. [cited by examiner]
Studer. Residue mutations and their impact on protein structure and function: detecting beneficial and pathogenic changes. Biochem. J. (2013) 449, 581-594. [cited by examiner]
Altschul, S., et al., “Basic local alignment search tool,” J. Mol. Biol., 215: 403-410 (1990). [cited by applicant]
Altschul, S.F., et al., “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” Nucleic Acids Res., 25(17):3389-3402 (1997). [cited by applicant]
Beaucage, S.L., et al., “Deoxynucleoside phosphoamidites—A new class of key intermediates for deoxypolynucleotide synthesis,” Tetrahedron Letters, 22(20):1859-62 (1981). [cited by applicant]
Botstein, D., et al., “Strategies and applications of in vitro mutagenesis,” Science, 229(4719):1193-1201 [1985]. [cited by applicant]
Carter, P., “Site-directed mutagenesis,” Biochem. J., 237:1-7 (1986). [cited by applicant]
Christians, F.C., et al., “Directed evolution of thymidine kinase for AZT phosphorylation using DNA family shuffling,” Nat. Biotechnol., 17:259-264 (1999). [cited by applicant]
Crameri, A., et al., “DNA shuffling of a family of genes from diverse species accelerates directed evolution”, Nature, 391:288-291 (1998). [cited by applicant]
Crameri, A., et al., “Improved green fluorescent protein by molecular evolution using DNA shuffling,” Nat. Biotechnol., 14(3):315-319 (1996). [cited by applicant]
Crameri, A., et al., “Molecular evolution of an arsenate detoxification pathway by DNA shuffling,” Nat. Biotechnol., 15(5):436-438 (1997). [cited by applicant]
Dale, S.J., et al., “Oligonucleotide-directed random mutagenesis using the phosphorothioate method,” Methods Mol. Biol., 57:369-74 (1996). [cited by applicant]
De Boer, H.A., et al., “The tac promoter: A functional hybrid derived from the trp and lac promoters,” Proc. Natl Acad. Sci. USA, 80: 21-25 (1983). [cited by applicant]
Ehrlich, S.D., “DNA cloning in Bacillus subtilis,” Proc Natl Acad Sci. USA, 75:1433 (1978). [cited by applicant]
Guo, Z., et al., “3'-End-Forming Signals of Yeast mRNA,” Mol. Cell. Biol., 15(11):5983-5990 [1995]. [cited by applicant]
Henikoff, S., et al., “Amino acid substitution matrices from protein blocks,” Proc. Natl. Acad. Sci. USA, 89:10915-10919 [1992]. [cited by applicant]
Koszelewski, D., et al., “Immobilization of omega-transaminases by encapsulation in a sol-gel/celite matrix,” Journal of Molecular Catalysis B: Enzymatic, 63: 39-44 (2010). [cited by applicant]
Kramer, B., et al., “Different base/base mismatches are corrected with different efficiencies by the methyl-directed DNA mismatch-repair system of [cited by applicant]
Lathe, R., et al., “Plasmid and bacteriophage vectors for excision of intact inserts,” Gene, 57:193-201 (1987). [cited by applicant]
Ling, M., et al., “Approaches to DNA Mutagenesis:An Overview,” Anal. Biochem., 254:157-78 (1997). [cited by applicant]
Martin, A.R., et al., “Characterization of free and immobilized (S)-aminotransferase for acetophenone production,” Applied Microbiology and Biotechnology, 76(4): 843-851 (2007). [cited by applicant]
Mateo, C., et al., “Epoxy sepabeads: a novel epoxy support for stabilization of industrial enzymes via very intense multipoint covalent attachment,” Biotechnology Progress 18(3):629-34 (2002). [cited by applicant]
Matthes, H.W.D., et al., “Simultaneous rapid chemical synthesis of over one hundred oligonucleotides on a microscale,” EMBO J., 3(4):801-05 (1984). [cited by applicant]
Minshull, J., et al., “Protein evolution by molecular breeding,” Curr. Op. Chem. Biol., 3(3):284-290 (1999). [cited by applicant]
Needleman, S., et al., “A general method applicable to the search for similarities in the amino acid sequence of two proteins,” J. Mol. Biol. 48:443-453 (1970). [cited by applicant]
Pearson, W.R., “Improved tools for biological sequence comparison,” Proc. Nat'l. Acad. Sci. USA, 85:2444-2448 (1988). [cited by applicant]
Rauwerdink, A., et al., “How the Same Core Catalytic Machinery Catalyzes 17 Different Reactions: the Serine-Histidine-Aspartate Catalytic Triad of α/β-Hydrolase Fold Enzymes,” ACS Cat., 5: 6153-6176 [2015]. [cited by applicant]
Romanos, M.A., et al., “Foreign gene expression in yeast: a review,” Yeast 8:423-488 [1992]. [cited by applicant]
Simonen, M., et al., “Protein Secretion in [cited by applicant]
Smith, M., “In vitro mutagenesis,” Ann. Rev. Genet., 19:423-462 (1985). [cited by applicant]
Smith, T., et al., “Comparison of Biosequences,” Adv. Appl. Math, 2:482-489 (1981). [cited by applicant]
Stemmer, W., “DNA Shuffling by Random Fragmentation and Reassembly: In vitro Recombination for Molecular Evolution,” Proc. Natl. Acad. Sci. USA, 91:10747-10751 (1994). [cited by applicant]
Stemmer, W.P.C., “Rapid evolution of a protein in vitro by DNA shuffling”, Nature, 370:389-391 (1994). [cited by applicant]
Straathof, A.J.J., et al., “The enantiomeric ratio: origin, determination and prediction,” Enzyme Microbio Technol, 21:559-571 [1997]. [cited by applicant]
Truppo, M.D., et al., “Development of an Improved Immobilized CAL-B for the Enzymatic Resolution of a Key Intermediate to Odanacatib,” Organic Process Research & Development, 15:1033-1035 (2011). [cited by applicant]
Varma, R., et al., “Lipase Catalysed Enantioselective Amidation of α-phenylethylamine,” Asian J. Biochem., 2(4): 279-283 [2007]. [cited by applicant]
Villa-Komaroff, L., et al., “A bacterial clone synthesizing proinsulin,” Proc. Natl Acad. Sci. USA, 75:3727-3731 (1978). [cited by applicant]
Wells, J.A., et al., “Cassette mutagenesis: an efficient method for generation of multiple mutations at defined sites,” Gene, 34:315-323 (1985). [cited by applicant]
Yi, S., et al., “Covalent immobilization of omega-transaminase from Vibrio fluvialis JS17 on chitosan beads,” Process Biochemistry 42(5): 895-898 (2007). [cited by applicant]
Zhang, J-H., et al., “Directed evolution of a fucosidase from a galactosidase by DNA shuffling and screening ,” Proc. Nat. Acad. Sci., U.S.A., 94:4504-4509 (1997). [cited by applicant]
Genbank accession No. WP_011038606.1 dated Sep. 11, 2023. [cited by applicant]
UniProt Accession No. A0A0H2X591 dated Sep. 16, 2015. [cited by applicant]
GenBank Accession No. RJU12542.1, dated Oct. 1, 2018. [cited by applicant]
Vorholter, F., et al., “The genome of Xanthomonas campestris pv. campestris B100 and its use for the reconstruction of metabolic pathways involved in xanthan biosynthesis,” J. Biotechnology, 134(1-2):33-45 [2008]. [cited by applicant]
UniProt Accession No. Q8P533 dated Oct. 1, 2002. [cited by applicant]
UniProt Accession No. B0RNI6 dated Apr. 8, 2008. [cited by applicant]