Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

A technology of glucosidase and glucan, applied in oligosaccharide, hydrolase, glycosylase, etc., can solve problems such as low-rate substrate concentration

Pending Publication Date: 2017-02-22
NUTRITION & BIOSCIENCE USA 4 INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this enzymatic activity occurs at a lower rate and in more dilute Substrate concentration

Method used

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  • Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes
  • Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes
  • Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0292] Preparation of Syrup by Polymerizing Sucrose

[0293] This example discloses a general approach to producing a mixture of soluble sugars by polymerizing sucrose using the gtf enzyme in a glucan synthesis reaction. Specifically, a filtrate of the dextran synthesis reaction was prepared, which was then concentrated into a syrup.

[0294] Sucrose (3000 g) was added to a clean 5-gallon polyethylene bucket. Water (18.1L) and Fermasure TM (10 mL) was added to the barrel, and by adding 5 vol% NaOH and 5 vol% H 2 SO 4 Adjust the pH to 7.0. The final volume was about 20 L and the initial concentration of sucrose was 152.5 g / L as determined by HPLC. Dextran polymerization was initiated by adding 0.3% by volume of crude gtfase (SEQ ID NO: 3) extract prepared as described in the General Methods section. The extract contained approximately 2.9 mg / mL protein. Agitation was provided to the reaction solution using an overhead mechanical motor equipped with a glass shaft and PT...

Embodiment 2

[0301] Effects of enzymes on the hydrolysis of sugars in the filtrate of dextran synthesis reactions

[0302] This embodiment measures various glucoamylase (EC 3.2.1.3), transglucosidase (EC 2.4.1.24), β-glucosidase (EC 3.2.1.21), α-amylase (EC 3.2.1.1 ) and glucosidase (EC 3.2.1) with the aim of reducing the concentration of leuconostoc and / or oligosaccharide by-products in the concentrated filtrate of the glucan synthesis reaction. Certain enzymes such as DIAZYME RDF ULTRA, transglucosidase (EC 2.4.1.24) and glucoamylase (EC 3.2.1.3), which are alpha-glucosidases, were found to be particularly effective in reducing the amount of these by-products, resulting in There was a corresponding increase in monosaccharides (glucose and fructose) in the treated filtrate.

[0303] According to the procedure outline of Example 1, the filtrate of the dextran synthesis reaction was first prepared and concentrated into a syrup. The composition of the concentrated filtrate is provided in...

Embodiment 3

[0330] Comparison of bond distributions of dextran reaction filtrate fractions before and after enzymatic hydrolysis

[0331] This example measures the effect of transglucosidase (EC 2.4.1.24) and β-glucosidase (EC 3.2.1.21) on the leuconostoc and oligosaccharide by-products present in the concentrated filtrate of the glucan synthesis reaction hydrolytic activity. Transglucosidase was found to reduce the amount of these by-products, resulting in a corresponding increase in the monosaccharides (glucose and fructose) in the treated filtrate.

[0332] The oligosaccharide by-products present in the filtrates of the above dextran synthesis reactions contained >90% glucose-glucose linkages as determined by NMR (general method). Of the glucose-glucose bonds, about 78% represent α-1,3 bonds and about 22% represent α-1,6 bonds.

[0333] NMR was used to determine the bond character of the material produced in Example 2.11 above after hydrolysis. Such as figure 1 As shown, the pea...

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Abstract

A method is disclosed for hydro!yzing an alpha-1,5 giucosyl-fructose linkage in a saccharide (disaccharide or oligosaccharide) such as leucrose. This method comprises contacting the saccharide with an alpha-glucosidase enzyme such as transgiucosidase or glucoamylase under suitable conditions, during which contacting step the enzyme hydrolyzes at least one alpha-1,5 glucosyl-fructose linkage of the saccharide. This method is useful for reducing the amount of leucrose in a filtrate isolated from a glucan synthesis reaction, for example.

Description

[0001] This application claims U.S. provisional applications 61 / 945,233 (filed February 27, 2014), 61 / 945,241 (filed February 27, 2014), 62 / 004,290 (filed May 29, 2014), 62 / 004,308 (filed 2014 29 May 2014), 62 / 004,312 (filed 29 May 2014), 62 / 004,300 (filed 29 May 2014), 62 / 004,314 (filed 29 May 2014) and 62 / 004,305 (filed May 29, 2014), the entire contents of all these applications are incorporated herein by reference. technical field [0002] The present invention is in the field of enzymatic hydrolysis of smaller carbohydrate polymers. In particular, the invention relates to the hydrolysis of disaccharides and oligosaccharides comprising one or more α-1,5 glucosyl-fructose linkages with alpha-glucosidases. [0003] References to Sequence Listings Submitted Electronically [0004] Electronically submit the official text of the sequence listing as a sequence listing in ASCII format via EFS-Web, the file name is CL6115USNP_SequenceListing_ST25.txt, the date of creation is Fe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/16C12P19/12C12P19/08C12P19/04C12P19/18
CPCC12P19/02C12P19/04C12P19/08C12P19/12C12P19/16C12P19/18C12Y302/0102C12Y204/01005C12Y204/01024C12Y302/01003C12Y302/0107C12Y302/01115C07H3/06C12Y302/01084C12N9/1051C12N9/2402C12N9/2428
Inventor K.D.纳盖E.C.哈戈J.K.舍蒂S.M.亨内塞R.迪科斯莫L.华R.拉米雷兹Z.汤Z.于
Owner NUTRITION & BIOSCIENCE USA 4 INC
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