Construction of a thermostable pullulanase producing bacillus strain and its fermentation studies

CHEN Chao; LIU Xiao-han; YU Feng; JI Ming-hua; SHI Ji-ping; SUN Jun-song

doi:10.13386/j.issn1002-0306.2015.20.037

CHEN Chao, LIU Xiao-han, YU Feng, JI Ming-hua, SHI Ji-ping, SUN Jun-song. Construction of a thermostable pullulanase producing bacillus strain and its fermentation studies[J]. Science and Technology of Food Industry, 2015, (20): 214-219. DOI: 10.13386/j.issn1002-0306.2015.20.037

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Construction of a thermostable pullulanase producing bacillus strain and its fermentation studies

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A codon optimized thermostable pullunase gene(Bn pul B) was synthesized and cloned behind an artificial tandom promter Pga2. The synthetic cassette was used to generate a suicide plasmid p GE-BPB;transformation of the plasmid into Bacillus subtilis 168 led to a pullulanase producing recombinant strain CH-1with native npr E deleted. Initially,30.3 U/m L of extracellular pullulanase was detected by CH-1 on regular medium supplemented with glucose,up to 268 U/m L of enzyme was detected by fermentation at 32 ℃ in medium with initial p H at 6.2,using 45 g/L of sucrose as carbon source and 60 g/L of wheat bran plus bean pulp as nitrogen source.

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Construction of a thermostable pullulanase producing bacillus strain and its fermentation studies

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