The acyl-CoA binding protein affects Monascus pigment production in Monascus ruber CICC41233

The acyl-CoA binding protein affects Monascus pigment production in Monascus ruber CICC41233
复制标题

酰基辅酶 A 结合蛋白影响红曲红 CICC41233 中红曲色素的产生。

DOI:
10.1007/s13205-018-1147-9
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发表时间:
2018-02-09
期刊:
影响因子:
2.8
通讯作者:
Zeng, Bin
Zeng, Bin
中科院分区:
工程技术4区
文献类型:
--
作者:
Long, Chuannan;Liu, Mengmeng;Zeng, Bin

文献摘要

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The present study verified whether acyl-coenzyme A (acyl-CoA)-binding protein (ACBP) affected the production of Monascus pigments (MPs) in Monascus ruber CICC41233 (MrACBP). Phylogenetic analysis revealed that the cloned Mracbp gene, which encoded the MrACBP protein, exhibited the closest match (99% confidence level) to the gene from Penicilliopsis zonata. The MrACBP and maltose-binding protein (MBP) were simultaneously expressed in Escherichia coli Rosetta DE3 in the form of a fusion protein. The microscale thermophoresis binding assay revealed that the purified MBP-MrACBP exhibited a higher affinity for myristoyl-CoA (Kd = 88.16 nM) than for palmitoyl-CoA (Kd = 136.07 nM) and octanoyl-CoA (Kd = 270.9 nM). Further, the Mracbp gene was homologously overexpressed in M. ruber CICC41233, and a positive transformant M. ruber ACBP5 was isolated. The fatty acid myristic acid in M. ruber ACBP5 was lower than that in the parent strain M. ruber CICC41233. However, when compared with the parent strain, the production of total MPs, water-soluble pigment, and ethanol-soluble pigment in M. ruber ACBP5 increased by 11.67, 9.80, and 12.70%, respectively, after 6 days. The relative gene expression level, as determined by a quantitative real-time polymerase chain reaction analysis, of the key genes acbp, pks, mppr1, fasA, and fasB increased by 4.03-, 3.58-, 1.67-, 2.11-, and 2.62-fold after 6 days. These data demonstrate the binding preference of MrACBP for myristoyl-CoA, and its influence on MPs production.