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Rhizopus and uses thereof

A technology of Rhizopus and application method, applied in the field of industrial microorganisms, can solve the problems of insufficient optical purity of products, difficult product quality control, serious environmental pollution, etc. Effect

Inactive Publication Date: 2009-08-19
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problems of the chemical method are that the optical purity of the product is not high enough, the operation is complicated, the environment is seriously polluted, the product cost is high, and the product quality is difficult to control.

Method used

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  • Rhizopus and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] This embodiment illustrates the screening purification method of Rhizopus Rhizopus sp.B1219:

[0028] PDA solid plate medium formula: 1.0L potato extract, 20.0g glucose, 20.0g agar, 1L distilled water, natural pH.

[0029] PDA liquid medium formula: 1.0L of potato extract, 20.0g of glucose, 1L of distilled water, natural pH.

[0030] 1. Breeding of starting strains:

[0031] Accurately weigh 25g of the wine medicine (taken from Changshu Wangsi Restaurant Yellow Wine Factory) for the production of rice wine, add the wine medicine into a triangular flask filled with 225mL of sterile water under aseptic conditions, shake for 30min, and absorb 1mL of the wine medicine dilution For further gradient dilution, draw 1mL of diluents of distilled spirits of different gradients and place them on the solidified sterile PDA plate medium, and spread the diluents of each gradient concentration evenly on the entire surface of the plate with a sterile applicator stick, 28 Incubate at ...

Embodiment 2

[0038] This example illustrates the application of Rhizopus sp.B1219 in the fermentative production of phenyllactic acid.

[0039] 1. Preparation of Rhizopus sp.B1219 mycelium

[0040] The above-mentioned Rhizopus spores were inoculated in 300mL PDA liquid medium, cultured in a shake flask at 28°C at 200r / min for 72 hours, centrifuged at 3000r / min, and separated from the bacterial liquid to obtain mycelium.

[0041] The formula of the PDA liquid medium is: 1.0L of potato juice, 20g of glucose, and natural pH.

[0042] 2. Fermentation of Rhizopus sp.B1219 mycelium to produce phenyllactic acid

[0043] The mycelium was inserted into 100mL (containing 39% phenylpyruvate) substrate conversion solution according to the inoculum size of 20g, reacted at 35°C for 12h, then extracted with 2 times the volume of ethyl acetate, evaporated and concentrated under reduced pressure to obtain The crude product of phenyl lactic acid, the content of phenyl lactic acid in the conversion solutio...

Embodiment 3

[0049] This example illustrates the application of Rhizopus sp.B1219 in the fermentative production of phenyl lactic acid, and the operation method is the same as the small-scale operation in Example 2. This example is the pilot-scale scale-up of Example 2.

[0050] By the method for embodiment 2, utilize 5L fermentors (with aseptic physiological saline the slant spores are washed down, adjust spore concentration to 10 7 Individuals / mL, inoculated into 3L culture medium, inoculum size 10%, shaking culture at 28°C for 120h) to cultivate mycelia, and transform in a 5L reaction tank (600g of mycelium was inserted into 3L substrate transformation solution) Reaction, the pH value is automatically adjusted with inorganic acid and alkali, and the separation and extraction of the product are carried out according to the method of Example 2. The result is: 35 ℃, conversion reaction time 14 hours, phenyl lactic acid output is 0.55g / L.

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Abstract

The invention relates to a Rhizopus sp. B1219 whose preservation date is January 6th, 2009, and the full name of the preservation institution is China Center for Type Culture Collection, CCTCC for short, and the preservation registration number is CCTCC No: M 209007; the invention further relates to the application of Rhizopus sp. B1219 in producing phenyl-lactic acid by fermentation. The Rhizopus sp. B1219 in the invention is derived from Chinese yeast for producing yellow wine, having broad application range for natural environments such as temperature and pH value, and the rhizopus can be easily cultivated and stored, and has strong ability of inhibiting food spoilage organisms, thereby guaranteeing the safety of use.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a rhizopus Rhizopus sp. B1219 strain and its application in fermentative production of phenyl lactic acid. Background technique [0002] β-Phenyllactic acid, also known as 3-phenyllactic acid or 2-hydroxy-3-phenylpropionic acid (Phenyllactic acid, referred to as PLA), has a relative molecular mass of 166 and a melting point of 121-125°C. It is odorless and resistant to acids, It is thermally stable and will not be destroyed when heated at 121°C. It naturally exists in honey and is non-toxic to human or animal cells. The second carbon atom of PLA is a chiral carbon atom, so there are two enantiomers R-(+)-3-phenyllactic acid (D-(+)-3-phenyllactic acid), and S- (-)-3-Phenyllactic acid (L-(-)-3-phenyllactic acid). PLA is a new type of antibacterial substance with a broad antibacterial spectrum, and its antibacterial properties have attracted extensive...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/56C12R1/845
Inventor 王立梅韩旭梅艳珍郑丽雪齐斌
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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