Protease gene shuffling and expression in Pichia pastoris



Document title: Protease gene shuffling and expression in Pichia pastoris
Journal: Brazilian archives of biology and technology
Database: PERIÓDICA
System number: 000387697
ISSN: 1516-8913
Authors: 1
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3
Institutions: 1Henan Agricultural University, College of Animal Science and Veterinary Medicine, Zhengzhou, Henan. China
2Henan Delin Biological Product Co. Ltd., Xinxiang, Henan. China
3Henan Engineering and Technology Research Center of Feed Microbes, Zhoukou, Henan. China
Year:
Season: May-Jun
Volumen: 58
Number: 3
Pages: 337-342
Country: Brasil
Language: Inglés
Document type: Artículo
Approach: Experimental, aplicado
English abstract Four kinds of neutral and alkaline protease genes from Aspergillus oryzae and Bacillus subtilis were isolated and shuffled. The shuffled genes were selected, inserted into pGAPZαA plasmid and transformed into Escherichia coli. The gene which could express high-activity protease was selected by screening the sizes of transparent zones around the colonies on casein plates. After an ideal protease gene was selected, it was sequenced and then transformed into Pichia pastoris X33. The result showed that the base in 1022th position of shuffled protease gene was changed from thymine to cytosine, inferring that cysteine was changed to arginine in the mutant protease. After 48 h incubation for the transformed P. pastoris with the mutant or native protease genes, the mutant protease activity was 36.4% higher than the native protease (P<0.05). The optimal pH and temperature of the mutant protease were 6.5-8.0 and 30-70°C, respectively, which indicated better stability than the native protease (P<0.05)
Disciplines: Biología
Keyword: Genética,
Hongos,
Bioquímica,
Proteasas,
Expresión génica,
Caracterización química,
Pichia pastoris
Keyword: Biology,
Fungi,
Genetics,
Biochemistry,
Proteases,
Gene expression,
Chemical characterization,
Pichia pastoris
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