| Registro completo |
| Provedor de dados: |
BABT
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| País: |
Brazil
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| Título: |
Protease gene shuffling and expression in Pichia pastoris
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| Autores: |
Yang,Gang
Chang,Juan
Yin,Qingqiang
Wang,Erzhu
Zhu,Qun
Wang,Ping
Dang,Xiaowei
Lu,Fushan
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| Data: |
2015-06-01
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| Ano: |
2015
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| Palavras-chave: |
Protease gene
DNA shuffling
Pichia pastoris
Gene expression
Protease characterization
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| Resumo: |
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).
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000300337
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| Editor: |
Instituto de Tecnologia do Paraná - Tecpar
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| Relação: |
10.1590/S1516-8913201500040
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| Formato: |
text/html
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| Fonte: |
Brazilian Archives of Biology and Technology v.58 n.3 2015
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| Direitos: |
info:eu-repo/semantics/openAccess
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