A lipase from Sporisorium reilianum SRZ2(SRL)with 73%amino acid sequence identity to Candida antarctica lipase B(CALB)was cloned and overexpressed in Pichia pastoris.The recombinant SRL showed a preference for short-c...A lipase from Sporisorium reilianum SRZ2(SRL)with 73%amino acid sequence identity to Candida antarctica lipase B(CALB)was cloned and overexpressed in Pichia pastoris.The recombinant SRL showed a preference for short-chain p-nitrophenyl esters.It achieved maximum activity at pH 8.0 and 65°C for p-nitrophenyl hexanoate(C6)with Km and kcar/Km values of 0.14 mmol·L-1 and 1712 min 1.mmol·L-1 at 30℃,respec-tively.SRL displayed excellent thermostability and pH stability,retaining more than 79%of its initial activity after incubation at 60℃ for 72h and 75%atpH 3to 11 for 72 h.It also maintained most of its activity in the presence of inhibitors and detergents except sodium dodecyl sulfate,and it tolerated organic solvents.SRL was covalently immobilized and successfully used for ethyl hexanoate synthesis in cyclohexane or in a solvent-free system with a high conversion yield(>95%).Furthermore,high con-version yield was also achieved for the synthesis of various short-chain flavor esters when high substrate concentra-tions of2 mol.LI were applied.This study indicated that a.CALB-type lipase from S.reiliamum SRZ2 showed great potential in organic ester synthesis.展开更多
文摘A lipase from Sporisorium reilianum SRZ2(SRL)with 73%amino acid sequence identity to Candida antarctica lipase B(CALB)was cloned and overexpressed in Pichia pastoris.The recombinant SRL showed a preference for short-chain p-nitrophenyl esters.It achieved maximum activity at pH 8.0 and 65°C for p-nitrophenyl hexanoate(C6)with Km and kcar/Km values of 0.14 mmol·L-1 and 1712 min 1.mmol·L-1 at 30℃,respec-tively.SRL displayed excellent thermostability and pH stability,retaining more than 79%of its initial activity after incubation at 60℃ for 72h and 75%atpH 3to 11 for 72 h.It also maintained most of its activity in the presence of inhibitors and detergents except sodium dodecyl sulfate,and it tolerated organic solvents.SRL was covalently immobilized and successfully used for ethyl hexanoate synthesis in cyclohexane or in a solvent-free system with a high conversion yield(>95%).Furthermore,high con-version yield was also achieved for the synthesis of various short-chain flavor esters when high substrate concentra-tions of2 mol.LI were applied.This study indicated that a.CALB-type lipase from S.reiliamum SRZ2 showed great potential in organic ester synthesis.
