Granted Patent
B2
US 12,668,824 · App. 18/024,942 · Granted Jun 30, 2026
Engineered transaminase biocatalysts and methods for synthesizing chiral amines
Inventors:
Santhosh Sivaramakrishnan (Redwood City, CA); Ericka Bermudez (Aptos, CA); Nandhitha Subramanian (Cambridge, GB); David Entwistle (San Carlos, CA); Stephanie Marie Forget (Redwood City, CA)
Assignee:
Codexis, Inc.
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Abstract
The present disclosure provides engineered transaminase polypeptides for the production of amines, polynucleotides encoding the engineered transaminases, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases to prepare compounds useful in the production of active pharmaceutical agents.
Claims (33)
1 . An engineered transaminase comprising a polypeptide sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, sequence identity to SEQ ID NO: 6, or a functional fragment thereof, wherein said engineered transaminase comprises a 130Y substitution in said polypeptide sequence, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 6, and wherein the engineered transaminase has an increased activity as compared to the polypeptide of SEQ ID NO: 4 in converting 1-imidazo[1,2-a]pyridin-6-ylethanone (compound (2)) to (1S)-1-imidazo[1,2-a]pyridin-6-ylethanamine (compound (3)).
2 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, sequence identity to SEQ ID NO: 6, wherein said engineered transaminase further comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 13, 13/41/57/88/130/415/417, 13/41/57/89/97/417, 13/41/57/97/415/417, 13/41/57/97/415/417/443, 13/41/57/97/443, 13/41/57/417, 13/41/57/417, 13/41/88, 13/41/88/89, 13/41/88/89/97/415/443, 13/41/88/89/417, 13/41/88/97, 13/41/88/415/443, 13/41/88/443, 13/41/89/148/443, 13/41/89/417, 13/41/89/443, 13/41/97/417, 13/41/97/415, 13/41/97/415/417, 13/41/97/417, 13/41/97/417/443, 13/41/130/415/443, 13/41/415, 13/41/415/417, 13/41/415/443, 13/41/417, 13/41/417/443, 13/57/88/89/415/443, 13/57/88/97, 13/57/88/97/415/443, 13/57/88/415, 13/57/88/130/417/443, 13/57/88/415, 13/57/97/415/417/443, 13/57/97/417, 13/88/89/415/417, 13/88/89/415/417/443, 13/88/443, 13/88/415, 13/89/97/415/417, 13/89/97/417, 13/89/417, 13/97/148/415, 13/97/415, 13/97/415/417, 13/97/417, 13/415, 13/415/417, 13/417, 13/417/443, 13/415, 13/415/417, 13/415/417/443, 13/415/443, 13/417, 13/417/443, 13/443, 23/53/162/233/277/315/415/418/432, 23/53/315/417/418, 23/277/315/395/415/417/432, 23/277/395/417/418, 23/395/418, 23/418, 41, 41/57/88, 41/57/88/415/443, 41/57/148/415/417, 41/57/443, 41/57/415/417, 41/88/89/97/415, 41/88/89/415/417, 41/88/97/417, 41/88/415/417, 41/88/443, 41/97/148/415/417/443, 41/97/417, 41/97/417/443, 41/415, 41/415/417/443, 41/415/443, 41/415/443, 41/417, 41/417/443, 53/162, 53/162/395/417, 53/162/418/432, 53/233, 53/277/395, 53/277/395/417/418, 53/277/415/417, 57/88/97/415/443, 57/88/97/417, 57/88/97/417, 57/97/148/417/443, 57/417, 88, 88/89/417, 88/97/415/417/443, 88/417/443, 88/148/417/443, 88/415, 88/415/417, 88/415/417/443, 88/417, 89/97/415/417, 89/97/417, 89/443, 97, 97/148/415, 97/415, 97/415/417, 97/417, 415, 417, 443, 162/233/415/417, 162/395/415/417, 162/418, 233/315/415/417, 233/315/417, 277/395/415/418/432, 315, 315/415/418/432, 395/418, 415, 415/417, 415/417/418, 415/417/418/432, 415/417/443, 415/443, 417, and 443, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 6.
3 . The engineered polypeptide of claim 2 in which the residue differences at the residue positions 13, 41/57/415/419, 41/113/415, 53/57, 88, 88/89, 97/415, 148, 227, 260, 302, 355/415/419, 362, 417, and 443 are selected from 13A, 13E, 13G, 13K, 13S, 41V/57Y/415F/419D, 41V/113F/415F, 53M/57W, 88K, 88R, 88R/89L, 88V, 97A/415S, 148E, 148G, 260T, 302N, 355C/415S/419D, 362G, 417A, 417I, 417V, 443E, and 443M.
4 . The engineered polypeptide of claim 1 in which the amino acid sequence further comprises a combination of residue differences as compared to SEQ ID NO: 6 selected from:
T13A;
T13E;
T13G;
T13K;
T13S;
I41V, F57Y, R415F, and Q419D;
I41V, V113F, and R415F;
N53M and F57W;
L88K;
L88R;
L88R and M89L;
L88V;
S97A and R415S;
Q148E;
Q148G;
C260T;
E302N;
R355C, R415S, and Q419D;
H362G;
L417A;
L417I;
L417V;
K443E; and
K443M.
5 . The engineered polypeptide of claim 1 in which the transaminase has at least 1.2 fold increased activity as compared to the polypeptide of SEQ ID NO: 4 in converting 1-imidazo[1,2-a]pyridin-6-ylethanone (compound (2)) to (1S)-1-imidazo[1,2-a]pyridin-6-ylethanamine (compound (3)).
6 . The engineered polypeptide of claim 1 in which further the transaminase has increased enantioselectivity as compared to the polypeptide of SEQ ID NO: 4 in converting 1-imidazo[1,2-a]pyridin-6-ylethanone (compound (2)) to (1S)-1-imidazo[1,2-a]pyridin-6-ylethanamine (compound (3)).
7 . The engineered polypeptide of claim 1 in which the amino acid sequence comprises a sequence selected from SEQ ID NO: 64, 66, 68, 76, 84, 88, 94, 110, 112, 118, 120, 124, 126, 128, 132, 138, 144, 150, 170, 182, 202, 206, 224, 226, 240, 246, 274, 276, 282, 294, 296, 302, 310, 312, 332, 336, and 338.
8 . The engineered polypeptide of claim 1 in which the polypeptide is immobilized on a solid support.
9 . The engineered polypeptide of claim 8 in which the solid support is a bead or resin comprising polymethacrylate with epoxide functional groups, polymethacrylate with amino epoxide functional groups, styrene/DVB copolymer or polymethacrylate with octadecyl functional groups.
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: SIVARAMAKRISHNAN, SANTHOSH; BERMUDEZ, ERICKA; SUBRAMANIAN, NANDHITHA; ENTWISTLE, DAVID; FORGET, STEPHANIE
To: CODEXIS, INC.
Reel/Frame 066424/0230 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Feb 8, 2024
From: SIVARAMAKRISHNAN, SANTHOSH; BERMUDEZ, ERICKA; ENTWISTLE, DAVID; FORGET, STEPHANIE MARIE; SUBRAMANIAN, NANDHITHA
To: CODEXIS, INC.
Reel/Frame 066417/0599 →
Continuity (2)
Provisional Application
63084166
· Sep 28, 2020
References Cited (262)
US 4518692A
· Rozzell
· 1985
[cited by applicant]
US 4600692A
· Wood et al.
· 1986
[cited by applicant]
US 4826766A
· Rozzell
· 1989
[cited by applicant]
US 4950606A
· Stirling et al.
· 1990
[cited by applicant]
US 5169780A
· Stirling et al.
· 1992
[cited by applicant]
US 5300437A
· Stirling et al.
· 1994
[cited by applicant]
US 5316943A
· Kidman et al.
· 1994
[cited by applicant]
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 6197558B1
· Fotheringham
· 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
· Del Cardayre 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 et al.
· 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 8470564B2
· Dhawan 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 8852900B2
· Cabirol et al.
· 2014
[cited by applicant]
US 8932838B2
· Cabirol et al.
· 2015
[cited by applicant]
US 9029106B2
· Dhawan et al.
· 2015
[cited by applicant]
US 9139821B2
· Nazor
· 2015
[cited by examiner]
US 9228223B2
· Mundorff et al.
· 2016
[cited by applicant]
US 9388395B2
· Nazor
· 2016
[cited by examiner]
US 9512410B2
· Dhawan et al.
· 2016
[cited by applicant]
US 9593326B2
· Clark et al.
· 2017
[cited by applicant]
US 9708588B2
· Nazor
· 2017
[cited by examiner]
US 9944909B2
· Dhawan et al.
· 2018
[cited by applicant]
US 20060195947A1
· Davis et al.
· 2006
[cited by applicant]
US 20080213845A1
· Fotheringham et al.
· 2008
[cited by applicant]
US 20080220990A1
· Fox
· 2008
[cited by applicant]
US 20090312196A1
· Colbeck et al.
· 2009
[cited by applicant]
WO 9522625A1
· 1995
[cited by applicant]
WO 9533836A1
· 1995
[cited by applicant]
WO 9600787A1
· 1996
[cited by applicant]
WO 9700078A1
· 1997
[cited by applicant]
WO 9735966A1
· 1997
[cited by applicant]
WO 9827230A1
· 1998
[cited by applicant]
WO 0042561A2
· 2000
[cited by applicant]
WO 0175767A2
· 2001
[cited by applicant]
WO 2005005633A2
· 2005
[cited by applicant]
WO 2008127646A2
· 2008
[cited by applicant]
WO 2009152336A1
· 2009
[cited by applicant]
WO 2010144103A1
· 2010
[cited by applicant]
WO 2011017551A1
· 2011
[cited by applicant]
WO 2020053198A1
· 2020
[cited by applicant]
Chica et al. Curr Opin Biotechnol. Aug. 2005;16(4):378-84. (Year: 2005).
[cited by examiner]
Singh et al. Curr Protein Pept Sci. 2017, 18, 1-11 (Year: 2017).
[cited by examiner]
Bornscheuer et al. Curr Protoc Protein Sci. Nov. 2011;Chapter 26:Unit26.7. (Year: 2011).
[cited by examiner]
Yoshikuni et al. Curr Opin Chem Biol. Apr. 2007;11(2):233-9. (Year: 2007).
[cited by examiner]
Pavlidis, I.V., et al., “Identification of (S)-selective transaminases for the asymmetric synthesis of bulky chiral amines”, Nature Chemistry, 8(11): 1076-1082 [2016].
[cited by applicant]
Midelfort, K.S., et al., “Redesigning and characterizing the substrate specificity and activity of Vibrio fluvialis aminotransferase for the synthesis of imagabalin,” Protein Eng Des Sel, 26(1):25-33 [2013].
[cited by applicant]
Geneseq Accession No. AZQ80534, “ATP-dependent DNA helicase [
[cited by applicant]
Altschul et al., “Basic local alignment search tool,” J. Mol. Biol., 215: 403-410 [1990].
[cited by applicant]
Altschul et al.,“Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” Nucleic Acids Res.,25,17, 3389-3402 [1977].
[cited by applicant]
Baldino et al., “High-resolution in situ hybridization histochemistry,” Meth. Enzymol., 168:761-777 [1989].
[cited by applicant]
Beaucage et al., “Deoxynucleoside Phosphoramidite—A New Class of Key Intermediates for Deoxypolynucleotide Synthesis,” Tetrahedron Letters 22: 1859-69, 1981.
[cited by applicant]
Bolton et al., “A General Method for the Isolation of RNA Complementary to DNA,” Proc. Natl. Acad. Sci. USA 48,8:1390-1397 [1962].
[cited by applicant]
Botstein et al., “Strategies and applications of in vitro mutagenesis,” Science, 229, 4719:1193-1201 [1985].
[cited by applicant]
Breslauer et al., “Predicting DNA duplex stability from the base sequence,” Proc. Natl. Acad. Sci. USA, 83:3746-3750 [1986].
[cited by applicant]
Carter, “Site-directed mutagenesis,” Biochem. J., 237,1:1-7 [1986].
[cited by applicant]
Cho et al., “Redesigning the Substrate Specificityof v-Aminotransferase for the KineticResolution of Aliphatic Chiral Amines,” Biotechnol Bioeng. 99(2):275-84, 2008.
[cited by applicant]
Christians et al., “Directed evolution of thymidine kinase for AZT phosphorylation using DNA family shuffling,” Nat. Biotechnol., 17,3:259-264 [1999].
[cited by applicant]
Crameri et al., “DNA shuffling of a family of genes from diverse species accelerates directed evolution,” Nature, 391:288-291 [1998].
[cited by applicant]
Crameri et al., “Improved Green Fluorescent Protein by Molecular Evolution Using DNA Shuffling,” Nat. Biotechnol., 14:315-319 [1996].
[cited by applicant]
Crameri et al., “Molecular evolution of an arsenate detoxification pathway by DNA shuffling,” Nat. Biotechnol., 15:436-438 [1997].
[cited by applicant]
Dale et al., “Oligonucleotide-Directed Random Mutagenesis Using the Phosphorothioate Method,” Meth. Mol. Biol.,57:369-374 [1996].
[cited by applicant]
DeBoer 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]
Freier et al., “Improved free-energy parameters for predictions of RNA duplex stability,” Proc. Natl. Acad. Sci. USA 83,24:9373-9377 [1986].
[cited by applicant]
GenBank Acc. No. ABA47738.1, GI:76578263; dated Jan. 31, 2014.
[cited by applicant]
GenBank Acc. No. AEA39183.1, GI:327207066; dated Apr. 4, 2011.
[cited by applicant]
GenBank Acc. No. AM902716.1, GI:163258032; dated Feb. 27, 2015.
[cited by applicant]
GenBank Acc. No. YP_002257813.1, GI:207739420; dated Dec. 16, 2010.
[cited by applicant]
Guo et al., “3′-End-Forming Signals of Yeast mRNA,” Mol. Cell. Biol., 15, 11:5983-5990 [1995].
[cited by applicant]
Henaut et al., “Analysis and Predictions from
[cited by applicant]
Henikoff et al., “Amino acid substitution matrices from protein blocks,” Proc. Natl. Acad. Sci. USA 89, 22:10915 [1989].
[cited by applicant]
Höhne et al., “Biocatalytic Routes to Optically Active Amines,” Chem Cat Chem 1(1):42-51.
[cited by applicant]
Höhne et al., “Efficient Asymmetric Synthesis of Chiral Amines by Combining Transaminase and Pyruvate Decarboxylase,” ChemBioChem, 9:363-365, 2008.
[cited by applicant]
Hwang et al., “High-throughput screening method for the identification of active and enantioselective ω-transaminases,” Enzyme and Microbial Technology, 34:429-436, 2004.
[cited by applicant]
Introduction to Organic Chemistry,2d ed., Andrew Streitwieser and Clayton Heathcock (1981), pp. 169-171.
[cited by applicant]
Iwasaki et al., “Microbial synthesis of chiral amines by (R)-specific transamination with
[cited by applicant]
Iwasaki et al., “Microbial synthesis of (R)- and (S)-3,4-dimethoxyamphetamines throughstereoselective transamination,” Biotech. Lett. 25:1843-1846; 2003.
[cited by applicant]
Wetmur, J.G., “DNA Probes: Applications of the Principles of Nucleic Acid Hybridization,” Rev. Biochem. Mol. Biol. 26:227-259 [1991].
[cited by applicant]
Kierzek et al., “Polymer-supported RNA synthesis and its application to test the nearest-neighbor model for duplex stability,” Biochem., 25,24:7840-7846 [1986].
[cited by applicant]
Koszelewski et al., “Asymmetric Synthesis of Optically Pure Pharmacologically Relevant Amines Employing ω-Transaminases,” Adv. Syn. Catal. 350:2761-2766, 2008.
[cited by applicant]
Koszelewski et al., “Deracemization of Mexiletine Biocatalyzed by ω-Transaminases,” Org Lett. 11(21):4810-4812, 2009.
[cited by applicant]
Koszelewski et al., “Immobilization of ω-transaminases by encapsulation in a sol-gel/celite matrix,”J. Mol. Cat. B: Enzymatic, 63: 39-44 [2010].
[cited by applicant]
Kramer et al., “Different base/base mismatches are corrected with different efficiencies by the methyl-directed DNA mismatch-repair system of
[cited by applicant]
Ling et al., “Approaches to DNA Mutagenesis: An Overview,” Anal. Biochem., 254(2):157-178 [1997].
[cited by applicant]
Martin et al., “Characterization of free and immobilized(S)-aminotransferase for acetophenone production,” Appl. Microbiol. Biotechnol., 76(4): 843-851[2007].
[cited by applicant]
Mateo et al., “Epoxy sepabeads: a novel epoxy support for stabilization of industrial enzymes via very intense multipoint covalent attachment,” Biotechnol. Prog., 18(3):629-34 [2002].
[cited by applicant]
Matthes et al., “Simultaneous rapid chemical synthesis of over one hundred oligonucleotides on a microscale,” EMBO J., 3(4):801-05 (1984).
[cited by applicant]
McInerney, “GCUA: General Codon Usage Analysis,” Bioinformatics, 14(4), 372-373 [1998].
[cited by applicant]
Midelfort, KS et al., Chain A, Pyruvate transaminase. PDB: 4E3R_A. NCBI entry (online). National Center for Biotechnology Information. Jan. 2, 2013 [retrieved on Jan. 25, 2022]. Retrieved from the Internet URL:https://w…
[cited by applicant]
Minshull et al., “Protein evolution by molecular breeding,” Curr. Op. Chem. Biol., 3,3:284-290 [1999].
[cited by applicant]
Mount, Bioinformatics: Sequence and Genome Analysis, Chapter 8, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. [ 2001].
[cited by applicant]
Nakamura et al., “Codon usage tabulated from international DNA sequence databases: status for the year 2000,” Nucl. Acids Res., 28,1:292[2000].
[cited by applicant]
Needleman 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]
Organic Chemistry, 2d ed., Francis Carey (1992), pp. 328-331.
[cited by applicant]
Organic Chemistry, 5th Ed., John McMurry, Brooks/Cole Publishing (2000), pp. 398 and 408.
[cited by applicant]
Pearson et al., “Improved tools for biological sequence comparison,” Proc. Natl. Acad. Sci. USA 85:2444-2448 [1988].
[cited by applicant]
Romanos et al., “Foreign Gene Expression in Yeast: a Review,” Yeast 8:423-488 [1992].
[cited by applicant]
Rychlik et al., “Optimization of the annealing temperature for DNA amplification in vitro,” [published erratum appears in Nucleic Acids Res 1991;19:698]. Nucleic Acids Res., 8:6409-6412 [1990].
[cited by applicant]
Savolitinib, BioVision, dated Mar. 2020.
[cited by applicant]
Shin et al., “Comparison of the ω-Transaminases from Different Microorganisms and Application to Production of Chiral Amines,” Biosci. Biotechnol, Biochem. 65:1782-1788, 2001.
[cited by applicant]
Shin et al., “Exploring the Active Site of Amine: Pyruvate Aminotransferase on the Basis of the Substrate Structure-Reactivity Relationship: How the Enzyme Controls Substrate Specificity and Stereoselectivity,” J. Org. …
[cited by applicant]
Shin et al., “Kinetic resolution of chiral amines with ω-transaminase using an enzyme-membrane reactor,” Biotechnol Bioeng 73:179-187, 2001.
[cited by applicant]
Shin et al., “Purification, characterization, and molecular cloning of a novel amine:pyruvate transaminase from Vibrio fluvialis JS17,” Appl. Microbiol. Biotechnol. 61(5-6):463-471, 2003.
[cited by applicant]
Shin J et al., “Asymmetric synthesis of chiral amines with ω-transaminase,” Biotechnol. Bioeng. 65, 206-211, 1999.
[cited by applicant]
Shin J et al., “Kinetic resolution of α-methylbenzylamine with o-transaminase screened from soil microorganisms: Application of a biphasic system to overcome product inhibition,” Biotechnol. Bioeng. 55:348-358, 1997.
[cited by applicant]
Simonen et al., “Protein secretion in Bacillus species,” Microbiol. Rev., 57:109-137, 1993.
[cited by applicant]
Smith et al., “Comparison of biosequences,” Adv. Appl. Math., 2:482-489 [1981].
[cited by applicant]
Smith M., “In Vitro Mutagenesis,” Annual Review of Genetics, 19(1), 423-462 [1985].
[cited by applicant]
Stemmer, “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, “Rapid evolution of a protein in vitro by DNA shuffling,” Nature, 370:389-391 [1994].
[cited by applicant]
Stenico et al., “Codon usage in Caenorhabditis elegans: delineation of translational selection and mutational biases,” Nucleic Acids Research, 22(13), 2437-2446 [1994].
[cited by applicant]
Suggs et al., “In: Brown DD (ed) ICN-UCLA 23rd Symposium on Developmental Biology using Purified Genes,” pp. 683-693. Academic Press, New York [1981].
[cited by applicant]
Fasman, 1989, CRC Practical Handbook of Biochemistry and Molecular Biology, CRC Press, Boca Raton, FL, at pp. 3-70.
[cited by applicant]
Tiwari et al., “Prediction of probable genes by Fourier analysis of genomic sequences,” Comput. Appl. Biosci., 13:263-270 [1997].
[cited by applicant]
Truppo et al., “Development of an Improved Immobilized CAL-B for the Enzymatic Resolution of a Key Intermediate to Odanacatib,” Org. Proc. Res. Dev., 2011, 15, 1033-1035.
[cited by applicant]
Uberbacher et al., “Discovering and understanding genes in human DNA sequence using GRAIL,” Meth. Enzymol., 266:259-281 [1996].
[cited by applicant]
Van Ophem et al., “Substrate Inhibition of D-Amino Acid Transaminase and Protection by Salts and by Reduced Nicotinamide Adenine Dinucleotide: Isolation and Initial Characterization of a Pyridoxo Intermediate Related to…
[cited by applicant]
Villa-Komaroff L et al., “A bacterial clone synthesizing proinsulin,” Proceedings of the National Academy of Sciences, 75(8), 3727-3731, 1978.
[cited by applicant]
Wada et al., “Codon usage tabulated from the GenBank genetic sequence data,” Nucl. Acids Res., 20:2111-2118 [1992].
[cited by applicant]
Wells et al., “Cassette mutagenesis: an efficient method for generation of multiple mutations at defined sites,” Gene, 34,2-3:315-323 [1985].
[cited by applicant]
Wright, “The ‘effective number of codons’ used in a gene,” Gene, 87,1:23-29 [1990].
[cited by applicant]
Yi et al.,“Covalent immobilization of v-transaminase from Vibrio fluvialis JS17 on chitosan beads,” Proc. Biochem., 42(5): 895-898 [2007].
[cited by applicant]
Yonaha et al., “Distribution of ω-Amino Acid: Pyruvate Transaminase and Aminobutyrate: α-Ketoglutarate Transaminase in Microorganisms ,” Agric.Biol. Chem. 47 (10):2257-2265, 1983.
[cited by applicant]
Yun et al., “ω-Amino Acid:Pyruvate Transaminase from Alcaligenes denitrificans Y2k-2: a New Catalyst for Kinetic Resolution of β-Amino Acids and Amines,” Appl. Environ. Microbiol. 70:2529-2534, 2004.
[cited by applicant]
Yun et al., “Kinetic resolution of (R,S)-sec-butylamine using omega-transaminase from Vibrio fluvialis JS17 under reduced pressure,” Biotechnol. Bioeng. 87:772-778, 2004.
[cited by applicant]
Yun et al., “Use of Enrichment Culture for Directed Evolution of the Vibrio fluvialis JS17 ω-Transaminase, Which Is Resistant to Product Inhibition by Aliphatic Ketones,” Appl Environ Microbiol. 71(8):4220-4224, 2005.
[cited by applicant]
Yun K., et al., “Asymmetric Synthesis of (S)-α-Methylbenzylamine by Recombinant
[cited by applicant]
Zhang et al., “Directed evolution of a fucosidase from a galactosidase by DNA shuffling and screening,” Proc. Nat. Acad. Sci. U.S.A., 94,9:4504-4509 [1997].
[cited by applicant]
Zhu et al., “Biocatalytic asymmetric amination of carbonyl functional groups—a synthetic biology approach to organic chemistry,” Biotechnol J. 4(10):1420-31, [2009].
[cited by applicant]
Batzer, M.A., “Erratum: Structure and variability of recently inserted Alu family members”, Nucleic Acids Res 19:698-699 [1991].
[cited by applicant]