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A method for detecting the amount of Gibberella fujikura active bacteria in gibberellin fermentation broth

A technology of gibberella fujikura and fermentation liquid, which is applied in the field of fermentation, can solve the problems of complex components, difficulty in effective and accurate detection of biomass, etc., and achieve the effect of high detection sensitivity and simple and easy operation

Active Publication Date: 2020-12-11
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a kind of method that detects gibberellin gibberellin fermented liquid and the method for gibberellum active thallus of fujikura, to solve because gibberellin fermentation culture medium contains a large amount of insoluble matter, the composition is complicated and gibberellin fujikura itself filament It is difficult to effectively and accurately detect the biomass in the fermentation production process caused by the physical properties of fungi.

Method used

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  • A method for detecting the amount of Gibberella fujikura active bacteria in gibberellin fermentation broth
  • A method for detecting the amount of Gibberella fujikura active bacteria in gibberellin fermentation broth
  • A method for detecting the amount of Gibberella fujikura active bacteria in gibberellin fermentation broth

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Experimental program
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Effect test

Embodiment 1X

[0031] Embodiment 1 XTT-menadione reagent color method rapid determination of bacterial mass

[0032] 1. Preparation of culture medium:

[0033] 1) PDA formula: 200g of potatoes, 20g of glucose (sucrose), 20g of agar, 1000mL of water, pH 6.5 (solid medium can be prepared by adding agar, and the water is ultrapure water).

[0034] PDA configuration process: First, peel and remove the buds from the potatoes, cut them into pieces the size of corn, weigh 200g, add 1000mL of water, and boil for about 15 minutes until it becomes a potato paste. Filter with double-layer gauze to obtain the filtrate, take the filtrate to make up to 1000mL (if preparing solid medium, add agar and melt), add other nutritional molecular substances, adjust the pH value to about 6.5, keep it neutral, put it in test tubes by classification, and plug it Cotton plugs, bundled and sterilized, made into slant media, and subsequently checked for sterilization.

[0035] 2) YPD: yeast extract 10g / l; peptone 20g / ...

Embodiment 2

[0050] The mensuration of active thalline amount, gibberellin content and reducing sugar content in the fermentation process of embodiment 2

[0051] 1. Preparation of culture medium

[0052] 1) Seed liquid: corn flour 2%; sucrose 1.5%; peanut powder 1.5%; soybean meal 0.1%; K 2 PO 4 0.1%; MgSO 4 0.1%; pH natural sterilization conditions: 121°C, 30 minutes.

[0053] Configuration: After configuring all the reagents according to the ratio, put them into a sterilizing pot for sterilization, inoculate them in an ultra-clean bench after natural cooling, and place them in a shaker in a constant temperature culture room at 28°C for 48 hours.

[0054] 2) Fermentation liquid: corn flour 7.5%; rice flour 8.75%; soybean meal 0.5%; KH 2 PO 4 0.05%; MgSO 4 0.05%;K 2 SO 4 0.05%; pH natural sterilization conditions; 121°C, 30 minutes.

[0055] Configuration: After preparing the cornstarch and rice flour according to the formula, add 1 / 1000 amylase, bathe in a water bath at 80°C...

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Abstract

The invention discloses a method for detecting the active thallus amount of Fusarium fujikuroi in gibberellin fermentation broth, and belongs to the technical field of fermentation. The method comprises the steps: (1) the to-be-detected fermentation broth is taken and mixed with a pre-blended XTT-menadione color-producing reagent to form a reaction system, and to-be-detected broth is prepared through incubation; (2) the light density OD450 of the to-be-detected broth at 450 nm is measured through a spectrophotometer; and (3) the measured OD450 value is directly used for representing the activethallus amount of the Fusarium fujikuroi in the to-be-detected fermentation broth, or the dry cell weight in the to-be-detected fermentation broth is calculated according to a formula to represent the active thallus amount of the Fusarium fujikuroi. According to the detecting method, interference of the physical properties of insoluble substances, dead cell residues and filamentous thallus in thegibberellin fermentation broth to detection is greatly reduced, the thallus amount of the Fusarium fujikuroi in the gibberellin fermentation broth can be accurately reflected, operation is easy and convenient and easy to implement, cells do not need to be washed, the detection sensitivity is high, and repeatability is achieved.

Description

technical field [0001] The invention relates to the technical field of fermentation, in particular to a method for detecting the amount of gibberella fujikura active bacteria in gibberellin fermentation broth. Background technique [0002] As a kind of natural plant growth regulator, gibberellins have been verified for a long time and have been widely used in agriculture, forestry, horticulture and other productions, and have high economic value and commercial prospects. At present, the commercial gibberellins are mainly GA3, GA4 and GA7, among which GA3 is the most widely used gibberellin product at home and abroad. my country is a major producer and exporter of gibberellin, and the investment in scientific research related to gibberellin has lasted for more than half a century. Fusarium fujikuroi is the only strain used for industrial production of gibberellin. The titer of gibberellin at the current production level reaches about 2000 μg / ml. [0003] Effective biomass ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/31G01N21/78
Inventor 柳志强岑宇科林建光王俊优郑裕国
Owner ZHEJIANG UNIV OF TECH
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