Pichia kudriavzevii resistant to high temperature and stress and high in ethanol yield and application thereof

A technology of yeast and ethanol, which is applied in the field of Pichia pastoris, can solve the problems of inability to grow, produce ethanol, and inability to produce ethanol

Active Publication Date: 2020-02-28
GUANGXI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The most commonly used industrial strain for ethanol production in the prior art is Saccharomyces cerevisiae, but it cannot grow

Method used

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  • Pichia kudriavzevii resistant to high temperature and stress and high in ethanol yield and application thereof
  • Pichia kudriavzevii resistant to high temperature and stress and high in ethanol yield and application thereof
  • Pichia kudriavzevii resistant to high temperature and stress and high in ethanol yield and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The isolation and identification of embodiment 1 bacterial strain

[0026] 1.1 Sample collection

[0027] Samples of yeast strains collected from subtropical rotting fruit.

[0028] 1.2 Isolation of strains

[0029] The collected samples were inoculated into enrichment medium YPD (containing 10% glucose, 2% peptone and 1% yeast extract), and incubated at 42° C. for 24 hours. Take 100 μL of the culture and evenly spread it on a plate containing 2% glucose, 2% peptone and 2% agar, and incubate at a constant temperature of 42°C for 24 hours. A single colony on the plate was inoculated into a test tube containing 3 mL of YPD and cultured at 42° C. for 24 hours to measure ethanol production. The strain with the highest ethanol production was selected for identification.

[0030] 1.3 Extraction of strain DNA

[0031] Inoculate a single colony of the strain to be identified in 15 mL of YPD and culture for 24 hours, centrifuge at 3000g for 10 minutes, discard the supernata...

Embodiment 2

[0037] Embodiment 2 investigates the characteristic of isolated bacterial strain

[0038] The isolated strains were divided into single colonies on the plate, and a single colony was inoculated into 3mL YPD medium for overnight cultivation. Take the overnight culture to measure OD600 and dilute it to 0.2, inoculate 200 μL of culture with OD600 of 0.2 into a 10 mL fermentation tube, give a small amount of oxygen supply conditions, culture at different temperatures at 200 rpm for different times, take out 1 mL and use high performance liquid phase method to measure and calculate ethanol concentration percentage.

[0039] The growth curves of strains under different temperature, ethanol, formic acid, furfural, hydrogen peroxide and acetic acid stress were determined by turbidimetric method. The specific steps are: inoculate a single colony into 3mL YPD medium and cultivate overnight, take the overnight culture to measure OD600, dilute it to OD600 with YPD medium without additive...

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Abstract

The invention belongs to the field of biochemical engineering, and particularly relates to pichia kudriavzevii resistant to high temperature and stress and high in ethanol yield and application thereof in bioethanol production. Pichia kudriavzevii is collected under the number of CGMCC NO.18379. The strain has high-temperature resistance and can resist various fermentation inhibitors and stress agents such as 30mM furfural, 70mM acetic acid, 30mM formic acid, 30mM H2O2 and ethanol of 15%, thereby being an advantageous starting strain in industrial production of bioethanol.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and in particular relates to high-temperature-resistant, stress-resistant and high-yielding Pichia kudriazwii and its application. Background technique [0002] Bioethanol has been promoted as an alternative to fossil fuels due to its sustainability, renewability, and environmental friendliness. Using corn, sugarcane and cassava as raw materials for bioethanol is costly and directly competes with the food supply, while lignocellulosic materials are more suitable as raw materials for bioethanol production. [0003] The conversion of lignocellulosic biomass to ethanol mainly includes three steps: pretreatment, saccharification and fermentation. Pretreatment is to remove or change the structure of lignin as a physical barrier, which is a necessary condition for microbial saccharification and fermentation in the later stage. The pretreatment process is easy to form by-products, such as furfural,...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P7/10C12R1/84
CPCC12P7/10C12R2001/84C12N1/165Y02E50/10
Inventor 房文霞金城伊费纳伊阿玛瑞·杜布伊斯汪斌覃启剑廖桂艳
Owner GUANGXI ACAD OF SCI
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