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Rhodotorula mucilaginosa strain ZZR-1# and method using strain to produce cutinase

A technology of ZZR-1, Rhododendron, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve good application prospects, conducive to large-scale production, and reasonable production methods.

Inactive Publication Date: 2014-10-22
HENAN HENGRUIYUAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microorganisms that produce cutinase are also rare, mainly plant pathogenic fungi, and there is no report on cutinase-producing yeast so far

Method used

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  • Rhodotorula mucilaginosa strain ZZR-1# and method using strain to produce cutinase
  • Rhodotorula mucilaginosa strain ZZR-1# and method using strain to produce cutinase
  • Rhodotorula mucilaginosa strain ZZR-1# and method using strain to produce cutinase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A method for producing cutinase by adopting Rhodotorula gum ZZR-1#, the specific operation steps are:

[0047] Step 1. Slant cultivation: inoculate the Rhodotorula colloides ZZR-1# strain stored in the low-temperature refrigerator on the slant seed medium, and cultivate at 26° C. for 48 hours to obtain the strain cultured on the slant seed medium, and set aside;

[0048] Described slant seed culture medium is, every 1000 ml culture medium, add potato 100 grams, tryptone 1.0g, lactose 1.0g and agar 17.0g, surplus is water;

[0049] Step 2. Shake flask seed culture: take 2 inoculation loops of slant seed culture medium, inoculate it into a 250mL Erlenmeyer flask containing 100mL shake flask seed medium, and culture it on a shaking table at 27°C and 160r / min for 36h , to obtain the shake flask seed culture strain;

[0050] The formula of the shake flask seed culture medium is: for every 1000 ml culture medium, add lactose 1.5g, yeast powder 0.5g, sodium chloride 2.0g and ...

Embodiment 2

[0059] A method for producing cutinase by adopting Rhodotorula gum ZZR-1#, the specific operation steps are:

[0060] Step 1. Slant cultivation: inoculate the Rhodotorula colloides ZZR-1# strain stored in the low-temperature refrigerator on the slant seed medium, and cultivate it at 28° C. for 48 hours to obtain the strain cultured on the slant seed medium, and set aside;

[0061] Described slant seed culture medium is, every 1000 ml culture medium, add potato 100 grams, tryptone 5.0g, lactose 5.0g and agar 17.0g, surplus is water;

[0062] Step 2. Shake flask seed culture: take 2 inoculation loops of slant seed culture medium, inoculate in a 250mL Erlenmeyer flask with 100mL shake flask seed medium, and culture at 29°C, 160r / min shaker for 48h , to obtain the shake flask seed culture strain;

[0063] The formula of the shake flask seed culture medium is: for every 1000 ml culture medium, add lactose 15.0g, yeast powder 5.0g, sodium chloride 0.5g and magnesium sulfate 0.5g, t...

Embodiment 3

[0072] A strain of Rhodotorula japonicus ZZR-1#, classified as Rhodotorula japonicus ( Rhodotorula mucilaginosa ) has been deposited in the General Microorganism Center (CGMCC) of the China Committee for the Collection of Microbial Cultures, and the preservation number is CGMCC No. 8890.

[0073] A method for producing cutinase by adopting Rhodotorula gum ZZR-1#, the specific operation steps are:

[0074]Step 1. Slant cultivation: Inoculate Rhodotorula colloides ZZR-1# strains stored in a low-temperature refrigerator on the slant seed medium, and cultivate them at 27°C for 48 hours to obtain the strains cultured on the slant seed medium for future use;

[0075] Described slant seed culture medium is, every 1000 ml culture medium, add potato 100 grams, tryptone 3.0g, lactose 4.0g and agar 17.0g, surplus is water;

[0076] Step 2. Shake flask seed culture: take 2 inoculation loops of slant seed culture medium, inoculate it into a 250mL Erlenmeyer flask containing 100mL shake f...

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Abstract

A Rhodotorula mucilaginosa strain ZZR-1# is named as Rhodotorula mucilaginosa, is preserved in China General Microbiological Culture Collection Center, and has a preservation number of CGMCC No.8890. The method using Rhodotorula mucilaginosa ZZR-1# to produce cutinase comprises the following steps: carrying out slant culturing on the Rhodotorula mucilaginosa by a slant culture medium, carrying out shaking seed culture, enlarged culture, enlarged culture and fermentation tank culture, filtering the obtained fermentation liquid by a gauze, and centrifuging the obtained filtrate at 4DEG C under a rotating speed of 5500rpm for 30min to obtain a supernatant which is a crude cutinase liquid; and concentrating the crude cutinase liquid, and purifying to obtain cutinase. The method using the Rhodotorula mucilaginosa ZZR-1# to produce cutinase is designed against the characteristics of the strain, and has the advantages of reasonability, simplicity, low cost, high enzyme activity in the crude cutinase liquid, high cutinase output, and good application prospect.

Description

technical field [0001] The invention relates to a strain of rhodotorula japonicus, specifically a strain of rhodotorula japonicus ZZR-1# and a method for producing cutinase by the strain. Background technique [0002] Extracting natural active ingredient compounds from plants is to dissolve these compounds from the tissue structure of plants and separate them from solid tissues. The traditional natural product extraction methods include alkali cooking, acid cooking, and water or alcohol cooking, but these methods all involve high-temperature cooking, which may cause activity loss for highly active drugs with good efficacy; especially acidic or alkaline Under aggressive conditions, it is also possible to change the pharmacological structure of the compound, such as the hydrolysis of glycosidic bonds and ester bonds. Environmental pollution is obviously also a major drawback of traditional methods. The method of enzymatic degradation and extraction of natural medicinal comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12N9/18C12R1/645
Inventor 张学俊张文坤冉琴琴张萌萌李银环苏晓兰
Owner HENAN HENGRUIYUAN IND
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