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Method for producing low-temperature raw diastase by fermenting microorganisms

A technology of raw starch saccharification enzyme and microbial fermentation, applied in the direction of hydrolase and so on

Active Publication Date: 2013-03-20
贵州牂牁江实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on low-temperature raw starch glucoamylase and its industrialization, large-scale production and application have not been reported yet

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] (1) Culture medium preparation

[0013] ① Strain activation medium: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 20.0g, distilled water 1.0L, pH value 7.0, autoclaved at 121℃ for 30min.

[0014] ② Low-temperature acclimation medium for strains: raw starch 0.5-5.0g, peptone 0.1-2.0g, beef extract 0.1-2.0g, NH 4 NO 3 0.5~5.0g, K 2 HPO 4 0.5~5.0g, MgSO 4 .7H 2 O 0.1~2.0g, FeSO 4 .7H 2 O 0.01-0.5g, KCl 0.1-2.0g, agar 20.0g, distilled water 1.0L, pH value is natural, sterilized by high-pressure steam at 121℃ for 30min. The raw starch was sterilized by dry heat at 105°C for 2 hours, and then added aseptically at room temperature.

[0015] ③ Liquid seed medium: peptone 5.0-20.0g, yeast extract 5.0-20.0g, glucose 0.5-2.0g, K 2 HPO 4 1.0-5.0g, 1.0L tap water, natural pH, sterilized by high-pressure steam at 121℃ for 30min.

[0016] ④ Enzyme production medium: raw corn starch 30.0-100.0g, beef extract 10.0-20.0g, NaCl 3.0-5.0g, FeSO 4 ·7H 2 O 0.01~0.2g, K 2 HPO...

Embodiment 2

[0024] (1) Culture medium preparation

[0025] ① Strain activation medium: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 20.0g, distilled water 1.0L, pH value 7.0, autoclaved at 121℃ for 30min.

[0026] ② Low-temperature acclimation medium for strains: raw starch 0.5-5.0g, peptone 0.1-2.0g, beef extract 0.1-2.0g, NH 4 NO 3 0.5~5.0g, K 2 HPO 4 0.5~5.0g, MgSO 4 .7H 2 O 0.1~2.0g, FeSO 4 .7H 2 O 0.01-0.5g, KCl 0.1-2.0g, agar 20.0g, distilled water 1.0L, pH value is natural, sterilized by high-pressure steam at 121℃ for 30min. The raw starch was sterilized by dry heat at 105°C for 2 hours, and then added aseptically at room temperature.

[0027] ③ Liquid seed medium: peptone 5.0-20.0g, yeast extract 5.0-20.0g, glucose 0.5-2.0g, K 2 HPO 4 1.0-5.0g, 1.0L tap water, natural pH, sterilized by high-pressure steam at 121℃ for 30min.

[0028] ④ Enzyme production medium: raw corn starch 30.0-100.0g, beef extract 10.0-20.0g, NaCl 3.0-5.0g, FeSO 4 ·7H 2 O 0.01~0.2g, K 2 HPO...

Embodiment 3

[0036] (1) Culture medium preparation

[0037] ① Strain activation medium: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 20.0g, distilled water 1.0L, pH value 7.0, autoclaved at 121℃ for 30min.

[0038] ② Low-temperature acclimation medium for strains: raw starch 0.5-5.0g, peptone 0.1-2.0g, beef extract 0.1-2.0g, NH 4 NO 3 0.5~5.0g, K 2 HPO 4 0.5~5.0g, MgSO 4 .7H 2 O 0.1~2.0g, FeSO 4 .7H 2 O 0.01-0.5g, KCl 0.1-2.0g, agar 20.0g, distilled water 1.0L, pH value is natural, sterilized by high-pressure steam at 121℃ for 30min. The raw starch was sterilized by dry heat at 105°C for 2 hours, and then added aseptically at room temperature.

[0039] ③ Liquid seed medium: peptone 5.0-20.0g, yeast extract 5.0-20.0g, glucose 0.5-2.0g, K 2 HPO 4 1.0-5.0g, 1.0L tap water, natural pH, sterilized by high-pressure steam at 121℃ for 30min.

[0040] ④ Enzyme production medium: raw corn starch 30.0-100.0g, beef extract 10.0-20.0g, NaCl 3.0-5.0g, FeSO 4 ·7H 2 O 0.01~0.2g, K 2 HPO...

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Abstract

The invention discloses a method for producing low-temperature raw diastase by fermenting microorganisms. The method comprises the steps of: performing low-temperature domestication on microorganisms producing raw diastase step by step to ensure that the microorganisms grow well in low-temperature environment; performing amplification culture on raw-diastase-producing bacteria obtained after low-temperature domestication step by step at a temperature of between 14 and 20 DEG C, inoculating into a liquid fermentation culture medium according to the inoculation amount which is 3 to 9 percent of the volume of fermentation liquor, and culturing for 72 to 144h at a temperature of between 14 and 20 DEG C, namely finishing the production of the low-temperature raw diastase by fermenting the microorganisms; centrifuging the fermentation liquor at a rotating speed of between 4,000 and 8,000rpm to collect liquid, wherein the collected liquid is crude enzyme fluid; and further concentrating, separating and purifying the crude enzyme fluid according to different needs and different using objects to obtain enzyme preparations with different activities, purities and formulations.

Description

technical field [0001] The invention relates to the fields of microbiology, enzyme engineering, fermentation engineering, biochemistry and the like, in particular to a microbial fermentation production method of low-temperature raw starch glucoamylase. The low-temperature raw starch glucoamylase produced by the invention is mainly used in grain processing, food industry, brewing, fermentation, textile industry, pharmaceutical industry and other starch glucoamylase application industries. It can improve the utilization rate, conversion rate and productivity of raw materials, reduce production costs, and improve and enhance product quality. Background technique [0002] Raw starch glucoamylase refers to enzymes that can directly hydrolyze raw starch without cooking and gelatinization into glucose. F, Food and Fermentation Industry, 1988). As a biological agent, raw starch glucoamylase has been widely used in grain processing, food industry, brewing, fermentation, textile ind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24
Inventor 张庆芳迟乃玉孙子羽冯旭明
Owner 贵州牂牁江实业有限公司