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Method for producing low temperature cellulase using microbe fermentation

A low-temperature cellulase and microbial fermentation technology, applied in biochemical equipment and methods, microorganisms, microorganisms, etc., can solve the problems of large-scale production and application of low-temperature cellulase that have not been reported

Inactive Publication Date: 2006-07-26
迟乃玉 +1
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  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been more than 40 years of research on cellulase at home and abroad, it is mainly the study of medium-temperature cellulase; the research on low-temperature cellulase is in its infancy, mainly focusing on strain selection and optimization of enzyme production conditions, growth characteristics and enzymology. Nature, and basic research on the cloning of related genes (Wang Bing et al., Shandong Institute of Education Journal and Marine Science, 2003; Zeng Yinxin et al., High Technology Communication, 2005; You Yinwei, Microbiology Journal, 2005); low-temperature cellulase industrialization , Large-scale production and application have not been reported

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: 200.0g potato, 20.0g glucose, 20.0g agar, 1.0L distilled water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0014] ②Low-temperature acclimation medium for strains: 50.0-100.0g potato, 5.0-8.0g Xinhua filter paper powder, 2.0-6.0g bran, 20.0g agar, 1.0L distilled water, natural pH, autoclaved at 121℃ for 30min.

[0015] ③Liquid seed medium: rice straw powder 3.0~5.0g, peptone 0.5~1.0g, Tween800.1~0.3g, (NH 4 ) 2 SO 4 0.8~1.2g, KH 2 PO 4 1.2~1.8g, urea 0.2~0.4g, MgSO 4 0.2~0.3g, CaCl 2 0.1~0.2g, ZnSO 4 ·7H 2 O 1.2 mg, MnSO 4 ·H 2 O 1.6 mg, CoCl 2 1.8mg, 1.0L tap water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0016] ④ Enzyme production medium: rice straw powder 7.0-12.0g, bran 1.0-5.0g, Tween800.1-0.4g, (NH 4 ) 2 SO 4 1.0~1.5g, KH 2 PO 4 1.4~2.0g, urea 0.2~0.6g, MgSO 4 0.1~0.4g, CaCl 2 0.2~0.3g, ZnSO 4 ·7H 2 O 2.4 mg, MnSO 4 ·H 2 O 3...

Embodiment 2

[0024] (1) Culture medium preparation

[0025] ① Strain activation medium: 200.0g potato, 20.0g glucose, 20.0g agar, 1.0L distilled water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0026] ②Low-temperature acclimation medium for strains: 50.0-100.0g potato, 5.0-8.0g Xinhua filter paper powder, 2.0-6.0g bran, 20.0g agar, 1.0L distilled water, natural pH, autoclaved at 121℃ for 30min.

[0027] ③Liquid seed medium: rice straw powder 3.0~5.0g, peptone 0.5~1.0g, Tween800.1~0.3g, (NH 4 ) 2 SO 4 0.8~1.2g, KH 2 PO 4 1.2~1.8g, urea 0.2~0.4g, MgSO 4 0.2~0.3g, CaCl 2 0.1~0.2g, ZnSO 4 ·7H 2 O 1.2 mg, MnSO 4 ·H 2 O 1.6 mg, CoCl 2 1.8mg, 1.0L tap water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0028] ④ Enzyme production medium: rice straw powder 7.0-12.0g, bran 1.0-5.0g, Tween800.1-0.4g, (NH 4 ) 2 SO 4 1.0~1.5g, KH 2 PO 4 1.4~2.0g, urea 0.2~0.6g, MgSO 4 0.1~0.4g, CaCl 2 0.2~0.3g, ZnSO 4 ·7H 2 O 2.4 mg, MnSO4 ·H 2 O 3.2 ...

Embodiment 3

[0036] (1) Culture medium preparation

[0037] ① Strain activation medium: 200.0g potato, 20.0g glucose, 20.0g agar, 1.0L distilled water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0038] ②Low-temperature acclimation medium for strains: 50.0-100.0g potato, 5.0-8.0g Xinhua filter paper powder, 2.0-6.0g bran, 20.0g agar, 1.0L distilled water, natural pH, autoclaved at 121℃ for 30min.

[0039] ③Liquid seed medium: rice straw powder 3.0~5.0g, peptone 0.5~1.0g, Tween800.1~0.3g, (NH 4 ) 2 SO 4 0.8~1.2g, KH 2 PO 4 1.2~1.8g, urea 0.2~0.4g, MgSO 4 0.2~0.3g, CaCl 2 0.1~0.2g, ZnSO 4 ·7H 2 O 1.2 mg, MnSO 4 ·H 2 O 1.6 mg, CoCl 2 1.8mg, 1.0L tap water, natural pH, sterilized by high pressure steam at 121℃ for 30min.

[0040] ④ Enzyme production medium: rice straw powder 7.0-12.0g, bran 1.0-5.0g, Tween800.1-0.4g, (NH 4 ) 2 SO 4 1.0~1.5g, KH 2 PO 4 1.4~2.0g, urea 0.2~0.6g, MgSO 4 0.1~0.4g, CaCl 2 0.2~0.3g, ZnSO 4 ·7H 2 O 2.4 mg, MnSO 4 ·H 2 O 3....

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Abstract

This invention relates to a microbe ferment producing low-temperature cellulose method, which is to domesticate the microbe that produces cellulose gradually under low temperature, which makes the microbe grows better; culturing the domesticated cellulose under 10-16deg C by expanding gradually and then inoculating 3-9% cultured cellulose by ferment liquid volume in the liquid ferment culture medium, when culturing for 72-144h in 10-16deg C, that means the microbe ferment producing low-temperature cellulose is complete; the ferment liquid collects liquid in 4, 000-8, 000rpm by centrifugal separation, the collected liquid is the coarse enzyme liquid; According to the different users and demand, the coarse enzyme can be condensed, separated and purified to be prepared into the enzyme preparation with different activity and pure. '

Description

technical field [0001] The invention relates to the fields of microbiology, enzyme engineering, fermentation engineering, biochemistry and the like, in particular to a low-temperature cellulase microbial fermentation production method. The low-temperature cellulase produced by the invention is mainly used in feed industry, food processing, extraction of Chinese herbal medicine, papermaking and textile, crude oil extraction, printing, dyeing and washing, and other cellulase 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] Cellulase refers to the general term of a group of enzymes that can degrade cellulose to generate small molecular substances such as cellobiose and glucose. It is a compound induced enzyme, including a variety of hydrolytic enzymes (Zhang Qixian, Acta Microbiology, 1974). As a biological agent, cellul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N1/00
Inventor 迟乃玉张庆芳
Owner 迟乃玉
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