Isolation of laccase gene from Bacillus subtilis and analysis of its physicochemical properties

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• 作者：Sui-Shan Yang^a ; Zhen-Wang Liu^a ; Xiao-Ping Yi^a ; Ai-Lian Zhang^a ;   ; ^{zhangailian6@yahoo.com.cn} ; Tian-Yuan Zhang^b ; Jin-Xian Luo^b ; Ze-Hua Zhang^a ; Jin-Cheng Shen^a ; Hui-Xiang Yin^a ; Li-Ping Chen^a
• 关键词：Laccase ; Gene cloning ; Expression ; Enzyme activity ; Physicochemical property
• 刊名：Gene
• 出版年：2012
• 出版时间：1 January 2012
• 年：2012
• 卷：491
• 期：1
• 页码：49-52
• 全文大小：364 K

文摘

The present study reports the cloning and sequencing of lac2 from Bacillus subtilis. The gene is composed of 1542 bp and encodes a 514-amino acid protein. The gene has 86 % homology with a published laccase with GeneID 936023. The lac2 gene was deposited in GenBank as a new nucleotide sequence. This new sequence was cloned into the multiple cloning site of pPIC9K to generate pPIC9K-lac2, which was then transformed into Pichia pastoris GS115 via electroporation. The recombinant GS115 (pPIC9K-lac2) was grown initially in BMGY medium and transferred to BMMY to induce gene expression for 48 h. The recombinant Lac2 protein shows laccase activity with ¦Á-naphthol and guaiacol as substrates. The optimal pH is between 3.2 and 4.7, and the optimal temperature is 25 ¡ãC for enzyme reaction.