IP Library Granted Patent US 9,657,045
Granted Patent B2
US 9,657,045 · App. 14/346,410 · Granted May 23, 2017

Process for producing a particulate composition comprising crystalline trehalose dihydrate

Inventors: Takashi Shibuya (Okayama, JP); Seisuke Izawa (Okayama, JP)
Assignee: HAYASHIBARA CO., LTD.
C07H3/04C07H1/00C12N9/1074C12P19/12C12P19/14C12P19/18C12P19/20C12Y204/01019C13K13/007
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Quick Facts
Patent No.
US 9,657,045
App. No.
14/346,410
Granted
May 23, 2017
Kind
B2
Abstract

A process for enabling the production of a particulate composition containing crystalline trehalose dihydrate is provided. Including allowing an α-glycosyltrehalose-forming enzyme to act on liquefied starch derived from a microorganism of the genus Arthrobacter and a trehalose-releasing enzyme derived from a microorganism of the genus Arthrobacter along with a starch debranching enzyme and a cyclomaltodextrin glucanotransferase; allowing glucoamylase to act on the resulting mixture to obtain a saccharide solution containing α,α-trehalose; precipitating crystalline α,α-trehalose dihydrate from the above saccharide solution; collecting the precipitated crystalline α,α-trehalose dihydrate by a centrifuge; and ageing and drying the collected crystals. Cyclomaltodextrin glucanotransferase derived from a microorganism of the genus Paenibacillus or a mutant enzyme thereof is used to increase the α,α-trehalose content in the saccharide solution to over 86.0% by weight, on a dry solid basis, without passing through a fractionation step by column chromatography.

Claims (25)

1. A process for producing a particulate composition comprising crystalline α,α-trehalose dihydrate with an α,α-trehalose content of 98.0% by weight or more, on a dry solid basis, said process comprising the steps of:

allowing to act on liquefied starch an α-glycosyltrehalose-forming enzyme derived from a microorganism of the genus Arthrobacter and a trehalose-releasing enzyme derived from a microorganism of the genus Arthrobacter along with a starch debranching enzyme and a cyclomaltodextrin glucanotransferase;

allowing a glucoamylase to act on the resulting mixture to obtain a saccharide solution comprising α,α-trehalose;

precipitating crystalline α,α-trehalose dihydrate from said saccharide solution by a controlled cooling method or semi-controlled cooling method;

collecting the precipitated crystalline α,α-trehalose dihydrate by a centrifuge; and

ageing and drying the collected crystals to obtain a particulate composition comprising crystalline α,α-trehalose dihydrate;

wherein a natural or recombinant cyclomaltodextrin glucanotransferase derived from a microorganism of the genus Paenibacillus or a mutant enzyme thereof is used as said cyclomaltodextrin glucanotransferase for converting maltooligosaccharides having a glucose polymerization degree of four or lower into maltooligosaccharides having a glucose polymerization degree of five or more, by a disproportionation reaction, and to increase the α,α-trehalose content in said saccharide solution to over 86.0% by weight, on a dry solid basis, without passing through a fractionation step by column chromatography;

wherein said controlled cooling method is a cooling method where the liquid temperature “T” at the time “t” is basically expressed by the formula:

T=T 0 −( T 0 −T f )( t /τ) 3 ,

wherein “τ” is the operation time established for a crystallization step, “T 0 ” is the liquid temperature at the initiation of crystallization, and “T f ” is the targeted liquid temperature at the termination of crystallization;

wherein said semi-controlled cooling method is a cooling method, where the liquid temperature “T” is allowed to linearly or stepwisely decrease against the time “t” in order to keep the variation (T 0 −T m ) of the liquid temperature “T” at the point of “t=τ/2” to be at least 5% but less than 50% of the total temperature change (T 0 −T f ), wherein “T m ” is the liquid temperature at time “t=τ/2”; and

wherein said particulate composition has a degree of crystallinity for crystalline α,α-trehalose dihydrate of 90.0% or higher, when calculated based on its powder X-ray diffraction profile.

2. The process of claim 1 , wherein said microorganism of the genus Paenibacillus is of the species Paenibacillus illinoisensis, Paenibacillus pabuli , or Paenibacillus amylolyticus.

3. The process of claim 1 , wherein said cyclomaltodextrin glucanotransferase is an enzyme that has the following partial amino acid sequences of (a) to (d):

(a)

Gly-Ser-X 1 -Ala-Ser-Asp;

(b)

Lys-Thr-Ser-Ala-Val-Asn-Asn;

(c)

Lys-Met-Pro-Ser-Phe-Ser-Lys;

and

(d)

Val-Asn-Ser-Asn-X 2 -Tyr;

wherein X 1 means Ala or Ser and X 2 means Ala or Thr, respectively.

4. The process of claim 1 , wherein said cyclomaltodextrin glucanotransferase is the one that has an amino acid sequence of SEQ ID NO: 1, 2, 3, 12 or 13.

Assignments (2)
CHANGE OF NAME Recorded Mar 18, 2025
From: HAYASHIBARA CO., LTD.
To: NAGASE VIITA CO., LTD.
Reel/Frame 070538/0828 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 21, 2014
From: SHIBUYA, TAKASHI; IZAWA, SEISUKE
To: HAYASHIBARA CO., LTD.
Reel/Frame 032938/0501 →
Priority Claims (2)
JP 2011-206482 · Sep 21, 2011 · national
JP 2012-168474 · Jul 30, 2012 · national
Continuity (1)
Related Publication 20150040889A1 · Feb 12, 2015