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Method for producing high maltose syrup by utilizing mpeg-mal5000-β-amylase

A technology of high maltose syrup and amylase, applied in the direction of fermentation, etc., can solve the problems such as β-amylase rarely reported, and achieve the effect of shortening the enzymatic hydrolysis time, lowering the enzymatic hydrolysis temperature, and reducing the production cost

Active Publication Date: 2021-12-07
HENAN INST OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Polyethylene glycol is a good chemical modifier, which has been used in various enzymes such as phospholipase, protease, arginine kinase, oxalate decarboxylase, etc., but its application in β-amylase is rarely reported

Method used

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  • Method for producing high maltose syrup by utilizing mpeg-mal5000-β-amylase
  • Method for producing high maltose syrup by utilizing mpeg-mal5000-β-amylase
  • Method for producing high maltose syrup by utilizing mpeg-mal5000-β-amylase

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Embodiment Construction

[0024] 1. Preparation of mPEG-Mal5000-β-amylase

[0025] 1 Materials and methods

[0026] 1.1 Materials and Instruments

[0027] 1.1.1 Reagents

[0028] β-amylase: American Sigma-Aldrich Company, molecular weight 55.7kDa, specific enzyme activity 17,000 U / mg; mPEG-NHS5000, mPEG-NHS20000, mPEG-Ts5000, mPEG-Ts10000, mPEG-Ts20000 and mPEG-Mal5000, Nanocs, USA company, purity ≥95%. All other reagents were of domestic analytical grade.

[0029] 1.1.2 Instruments and equipment

[0030] UV-Vis spectrophotometer: WFJ7200 (Unocal Shanghai Instrument Co., Ltd.);

[0031] Constant temperature water bath: DZKW-4 (Beijing Zhongxing Weiye Instrument Co., Ltd.);

[0032] Oscillating constant temperature metal bath: TUS-200P (Shanghai Yiheng Technology Co., Ltd.).

[0033] 1.2.2 mPEG modified β-amylase reaction

[0034] Add 8.0 mL of disodium hydrogen phosphate-citrate buffer solution of a certain pH to the test tube, then add 1.0 mL of 125 μmol / L β-amylase enzyme solution and 1.0 mL ...

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Abstract

The invention discloses a method for producing high maltose syrup with mPEG-Mal5000-β-amylase: ①Mix starch and water into starch slurry, add high-temperature-resistant α-amylase, keep warm at 90°C for 3 minutes, and inactivate the enzyme; ②Inactivate the enzyme Add mPEG‑Mal5000‑β‑amylase to the final starch solution, after enzymatic hydrolysis, boil and centrifuge to obtain high maltose syrup. Compared with unmodified β-amylase, the application of mPEG-Mal5000-β-amylase to produce high maltose syrup, the amount of enzyme added was reduced from 200 U / g to 120 U / g, and the usage was reduced by 40%; the saccharification time was reduced from 28h Reduced to 16h; yield increased from 73.29% to 85.67%. For enzymatic hydrolysis of corn starch, the amount of enzyme added was reduced from 200 U / g to 100 U / g, and the usage was reduced by 50%; the saccharification time was reduced from 27h to 14h; the yield increased from 76.53% to 86.34%.

Description

technical field [0001] The invention belongs to the field of plant starch utilization, and in particular relates to a method for producing high maltose syrup by utilizing mPEG-Mal5000-β-amylase. Background technique [0002] β-amylase is one of the amylases, and widely exists in higher plants such as barley, wheat, sweet potato, soybean, and microorganisms such as Bacillus. According to reports, in recent years, the post-harvest starch processing ratio of sweet potatoes in China has reached 70%-80%. China discharges about 16.5 million tons of waste liquid every year due to the production of potato starch. If it is not treated in time, it will pollute the environment. β-amylase in sweet potato is a protein component in tubers next to sweet potato storage protein. The β-amylase extracted from sweet potato is the best source of plant β-amylase. However, in actual production and application, it is found that β-amylase in sweet potato is easily affected by environmental conditi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/12C12P19/22
CPCC12P19/12C12P19/22
Inventor 梁新红赵功玲孙俊良冉军舰焦凌霞张万利冯龙斐莫海珍张军合
Owner HENAN INST OF SCI & TECH
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