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Low energy N ~ into the L-Ile bacteria Mutation Breeding and fermentation conditions

Author: WangXingLi
Tutor: ChenLinHai;LiZongWei
School: Zhengzhou University
Course: Biophysical
Keywords: L- isoleucine Corynebacterium crenatum Ion implantation Growth ring method. Paper chromatography - spectrophotometry Ferment
CLC: TQ920.1
Type: Master's thesis
Year: 2007
Downloads: 75
Quote: 3
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Abstract


The ion implantation for microbial mutagenesis to obtain a higher mutation rate in the case of light damage and wider mutation spectrum , and to a certain extent to overcome the resistance of strains of traditional mutagenesis source saturation . Through to produce bacteria Corynebacterium crenatum of L- isoleucine (Corynebacterium crenatum) AS1.998 the N ion implantation combined with UV mutagenesis and ethionine resistance screening , screening the mutagenic strain N24. L- isoleucine production from 0.446g/100ml to 0.642g/100ml, 43.95% higher than the original strain , the mutant has a good genetic stability . The single factor and orthogonal experiment method of seed medium mutation the strains N24 shake flask fermentation , the fermentation medium optimized . Seed medium : 7% glucose , 1.8% corn steep liquor , peptone 0.5 % , KH 2 PO 4 0.1% MgSO 4 0.05 % , urea 0.4% (single off) . Fermentation medium : glucose 12%, (NH 4 ) 2 SO 4 6%, K 2 HPO 4 0.06%, corn steep liquor 1.0% , 1.0% bran hydrolyzate . The optimization seed and fermentation medium , the mutant N24 L- isoleucine production of 0.744g/100ml, 15.89% than the original medium 0.642g/100ml . Mutant strain N24 flask fermentation conditions optimized initial pH 7.2 , CaCO , 3 to adjust the pH, inoculum 10% , medium volume 25ml/500ml , speed 200rpm , 28 ℃ fermentation 72h, L - isoleucine production reached 0.775g/100ml, compared with the original culture conditions to increase by 4.17% . Mutant N24 optimized fermentation medium and fermentation conditions , L - isoleucine production reached 0.775g/100ml, compared with the original strain 0.446g/100ml 73.77% , and has a good genetic stability .

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