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Galactomyces and method of in-situ cellulose enzymolysis using same

An in-situ enzymatic hydrolysis and cellulose technology, which is applied in the field of enzymatic hydrolysis of cellulose raw materials, achieves high activity, less waste, and process economy

Active Publication Date: 2014-06-18
兰溪市市场开发服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on NMMO-tolerant cellulase-producing bacteria, so screening new enzyme sources of NMMO-tolerant cellulase is a top priority

Method used

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  • Galactomyces and method of in-situ cellulose enzymolysis using same

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Cellulase fermentation in the presence of NMMO:

[0027] Fill a 1 L shake flask with 250 mL medium ((NH 4 ) 2 SO 4 10 g / L, KH 2 PO 4 8 g / L, MgSO 4 ·7H 2 O 0.8 g / L, CaCl 2 2H 2 O 0.6 g / L, Tween-80 1.2 g / L, CMC-Na 20 g / L, bagasse 20 g / L), pre-cultured on a shaker at 160 rpm at 35 °C Galactomyces sp. CCZU11-1 48 h. Add 10% (w / w) NMMO and continue culturing for 192 h. The filter paper enzyme activity (FPA) and endonuclease activity (CMCase) of the obtained cellulase fermentation broth were 0.45 U / mg and 0.63 U / mg, respectively.

Embodiment 2

[0029] (1) Galactomyces Culture of sp. CCZU11-1:

[0030] Fill a 1 L shake flask with 250 mL medium ((NH 4 ) 2 SO 4 4 g / L, KH 2 PO 4 2 g / L, MgSO 4 ·7H 2 O 0.1 g / L, CaCl 2 2H 2 O 0.1 g / L, Tween-80 0.2 g / L, CMC-Na 10 g / L, bagasse 10 g / L), cultivated on a shaker at 160 rpm at 25 °C Galactomyces sp. CCZU11-1 120 h. The filter paper enzyme activity (FPA) and endonuclease activity (CMCase) of the obtained cellulase fermentation broth were 0.48 U / mg and 0.76 U / mg, respectively. The fermented liquid obtained is ultrafiltered with a PVDF100 ultrafiltration membrane to obtain active crude protein, and the active protein is further freeze-dried to obtain cellulase freeze-dried powder.

[0031] (2) NMMO / solvent system enzymatic cellulose raw material:

[0032] 2.0 g / L bagasse and microcrystalline cellulose were pre-dissolved with 85% (w / w) NMMO at 85 °C for 35 min, and acetic acid-sodium acetate buffer (pH=5.0) was added (50 mM) form the NMMO / solvent system, the NMMO conten...

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Abstract

The invention relates to galactomyces and a method of in-situ cellulose enzymolysis using the same, belonging to the field of microorganism technology, and relates to cellulase-producing bacteria galactomyces sp.CCZU11-1 newly separated from soil and tolerant to morpholine oxide-N-oxide (NMMO, N-methylmorpholine-oxide), wherein the collection number of the galactomyces sp.CCZU11-1 is CGMCC No.5561. The metabolized cellulase is tolerable to the NMMO and still shows relatively high activity when 25% (v / v) of NMMO is contained in the reaction system, in-situ efficient enzymolysis of cellulose can be realized, and the advantages of economic process, few three wastes and the like are realized. The method provided by the invention has simple technology; and compared with the existing 'two-step method' of cellulose processing with NMMO and regeneration enzymolysis, the method has the advantages of high efficiency, economic process, few three wastes and the like.

Description

technical field [0001] The invention discloses a cellulase-producing fungus Geotrichum newly isolated from soil Galactomyces The cultivation of sp. CCZU11-1 (preservation number is CGMCC No. 5561) and its method for efficient in-situ enzymolysis of cellulose raw materials in NMMO / solvent system belong to the field of microbial technology. Background technique [0002] With the increasing scarcity of non-renewable resources such as coal and petroleum and the increasing attention and attention to environmental pollution in various countries, it is no longer possible to convert renewable resources such as biomass cellulose into high calorific value energy, new industrial raw materials, and fine chemicals. Avoid becoming the trend of future industrial development. According to the US Department of Energy, renewable resources such as cellulose will be used in industrial production on a large scale, and chemical products from renewable resources such as cellulose will account for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P19/14C12R1/645
Inventor 何玉财夏东琴张跃金银唐红梅马翠鸾王利群王旭东蔡蕾陈晓婧龚婷
Owner 兰溪市市场开发服务有限公司