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Microorganism strain for generating pectinase and application thereof in tobacco

A microbial strain and pectinase technology, applied in the directions of microorganism-based methods, microorganisms, applications, etc., to achieve the effects of improving sensory quality, increasing the amount and concentration of smoke aroma, and reducing the content of tobacco pectin

Active Publication Date: 2013-07-10
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, studies have shown that some strains of Penicillium verruculosum can produce cellulase (Seonghun Kim and Chul Ho Kim.2012. Production of cellulase enzymes during the solid-state fermentation of empty palm fruit bunch fiber. Bioprocess Biosyst Eng, 35(1-2):61-67), unpublished reports on the pectinase-producing properties of this microbial strain and its application in tobacco

Method used

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  • Microorganism strain for generating pectinase and application thereof in tobacco
  • Microorganism strain for generating pectinase and application thereof in tobacco
  • Microorganism strain for generating pectinase and application thereof in tobacco

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Add 5mL of potato dextrose agar medium to a 20mL test tube to make a slant. Penicillium verruculosum The TS63-9 pectinase-producing strain was inoculated on a slant and cultured at 28°C for 48 hours, added 5 mL of sterile water, and oscillated to obtain a spore suspension. The spore suspension was inoculated into a 250mL Erlenmeyer flask containing 30mL fermentation medium at a ratio of 5% for fermentation. The culture temperature was 28° C., the rotation speed of the shaker was 200 rpm, and the culture time was 72 hours. The fermentation liquid was centrifuged at 4°C and 6000 rpm to obtain a supernatant which was the crude enzyme liquid. After testing, the pectinase activity of the obtained crude enzyme solution was 701.5 U·mL -1 .

[0043] The composition of the fermentation medium is (g / L): Tobacco stem powder 7.5, KH 2 PO 4 2.0, (NH 4 ) 2 SO 4 2.5, MgSO 4 · 7H 2 O 0.3, CaCl 2 0.3, Tween-80 1.0mL / L, pH 6.0.

[0044] The flue-cured tobacco leaves in Ch...

Embodiment 2

[0057] Add 5mL of potato dextrose agar medium to a 20mL test tube to make a slant. Penicillium verruculosum The TS63-9 pectinase-producing strain was inoculated on a slant and cultured at 28°C for 24 hours, added 5 mL of sterile water, and oscillated to obtain a spore suspension. The spore suspension was inoculated into a 250mL Erlenmeyer flask containing 30mL fermentation medium at a ratio of 5% for fermentation. The culture temperature was 28° C., the rotation speed of the shaker was 200 rpm, and the culture time was 120 h. The fermentation liquid was centrifuged at 4°C and 6000 rpm to obtain a supernatant which was the crude enzyme liquid. After testing, the pectinase activity of the obtained crude enzyme solution was 695.2 U·mL -1 .

[0058] The composition of the fermentation medium is (g / L): Tobacco stem powder 7.5, KH 2 PO 4 2.0, (NH 4 ) 2 SO 4 2.5, MgSO 4 ·7H 2 O 0.3, CaCl 2 0.3, Tween-80 1.5mL / L, pH 6.0.

[0059] The shredded stem samples were equilibr...

Embodiment 3

[0062] Add 5mL of potato dextrose agar medium to a 20mL test tube to make a slant. Penicillium verruculosum The TS63-9 pectinase-producing strain was inoculated on a slant and cultured at 28°C for 72 hours, added 5 mL of sterile water, and oscillated to obtain a spore suspension. The spore suspension was inoculated into a 250mL Erlenmeyer flask containing 30mL fermentation medium at a ratio of 5% for fermentation. The fermentation medium base uses orange peel powder as a carbon source, the culture temperature is 28°C, the shaker speed is 200rpm, and the culture time is 72h. The fermentation liquid was centrifuged at 4°C and 6000 rpm to obtain a supernatant which was the crude enzyme liquid. The pectinase activity of the obtained crude enzyme solution was determined to be 501.9 U·mL -1 .

[0063] The composition of the fermentation medium is (g / L): orange peel powder 5.0, KH 2 PO 4 2.0, (NH 4 ) 2 SO 4 1.5, MgSO 4 · 7H 2 O 0.3, CaCl 2 0.3, Tween-80 1.0mL / L, pH 6.0...

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Abstract

The invention relates to a microorganism strain for generating pectinase and an application of the microorganism strain in tobacco, and belongs to the field of microorganism fermentation and application. The microorganism strain is a Penicillium verruculosum TS63-9 strain which is preserved in China General Microbiological Culture Collection Center on October 31st, 2012 with the preservation number of CGMCC No. 6747. The ITSrRNA gene sequence of the TS63-9 strain is the main characteristic basis for distinguishing the strain. According to the microorganism strain, a crude fermentation enzyme liquid is obtained by liquid fermentation of tobacco stem powder or orange peel powder as a carbon source; the crude fermentation enzyme liquid is added into tobacco shreds or tobacco pieces according to an appropriate ratio, and then the pectin content of the tobacco is greatly reduced; after being treated, the tobacco is coordinated in chemical components, the smoking evaluation result shows that the fragrance is increased, and the concentration is increased, and the sensation quality is improved; and in addition, the crude fermentation enzyme liquid can also remarkably reduce the content of the pectin in the tobacco stems.

Description

Technical field: [0001] The invention relates to a pectinase-producing microbial strain and its application in tobacco, belonging to the field of microbial fermentation and application. Background technique: [0002] Pectin is a hydrophilic colloid that binds cell walls together between plant cells. It is a linear polymer formed by connecting D-galacturonic acid and methyl D-galacturonic acid with α-1,4 glycosidic bonds. . Pectinase is a general term for various enzymes that can decompose pectin, mainly produced by plants and microorganisms. Microbial pectinolytic enzymes: A review. Process Biochemistry, 40(9): 2931-2944. Bacteria, actinomycetes, yeast, and fungi can all secrete pectinase. [0003] Pectin, as a constituent of the cell wall of tobacco leaves, has an important influence on the quality of tobacco leaves and smoke components. Pectin plays a certain role in the hygroscopicity and elasticity of tobacco leaves, but when the content is too high, it will increas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/88C12N9/18C12N9/26A24B15/20C12R1/80
Inventor 宋纪真贺兆伟奚家勤范坚强邓宾玲韦建玉杨军郭建华高志强
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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