Screening, purification, and characterization of a novel organic solvent-tolerant esterase, Lip2, from Monascus purpureus strain M7

Screening, purification, and characterization of a novel organic solvent-tolerant esterase, Lip2, from Monascus purpureus strain M7
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紫红曲霉菌株 M7 中新型耐有机溶剂酯酶 Lip2 的筛选、纯化和表征

DOI:
10.1007/s00792-016-0907-x
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发表时间:
2017-01
期刊:
影响因子:
2.9
通讯作者:
Mu Li
Mu Li
中科院分区:
生物学3区
文献类型:
--
作者:
Li-Jing Kang;Zi-Tong Meng;Chen Hu;Yan Zhang;Hai-Lun Guo;Qing Li;Mu Li

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耐有机溶剂的酯酶在化工和制药工业中被证明是优良的生物催化剂。从丝状真菌Monastyrpurpureus M7中分离到一个新的耐有机溶剂酯酶基因lip 2。序列分析表明,lip 2在活性中心附近有一个保守的“GDSL”基序。多序列比对和系统发育分析表明,Lip 2显示了两个独特的氨基酸序列基序,清楚地将其与任何其他先前描述的脂肪酶家族分开。在20%甲醇和乙醇中孵育3小时后,Lip 2分别显示190%和180%的残留活性。孵育1天后,在20%(v/v)亲水性有机溶剂中保留99-110%的相对活性。该酯酶在40 °C下显示出最佳活性,并且在60 °C下保持约70%的最大活性。该酶在pH 5-11的范围内也显示出超过50%的残余活性。在大多数金属离子或添加剂存在下,Lip 2保留了大部分活性。Lip 2的这些独特性质使其成为工业过程的生物催化剂。
Organic solvent-tolerant esterases are proven to be excellent biocatalysts in chemical and pharmaceutical industries. A novel organic solvent-tolerant esterase gene, lip2, was isolated from filamentous fungi Monascus purpureus M7. The sequence analysis suggested that lip2 has a conserved "GDSL" motif near the active center. The multiple-sequence alignment and phylogenetic analysis revealed that Lip2 displayed two unique amino-acid sequence motifs that clearly separate it from any other previously described lipase family. After incubation in 20% methanol and ethanol for 3 h, the Lip2 displayed 190 and 180% residual activities, respectively. It retained 99-110% relative activity in 20% (v/v) hydrophilic organic solvents after incubation for 1 day. This esterase showed optimal activity at 40 °C and retained about 70% maximal activity at 60 °C. The enzyme also displayed more than 50% residual activity over a range of pH 5-11. In the presence of most of metal ions or additives, Lip2 retained most of the activity. These unique properties of Lip2 make it a promising as biocatalyst for industrial processes.
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