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- Title
The acyl-CoA binding protein affects <italic>Monascus</italic> pigment production in <italic>Monascus ruber</italic> CICC41233.
- Authors
Long, Chuannan; Liu, Mengmeng; Chen, Xia; Wang, Xiaofang; Ai, Mingqiang; Cui, Jingjing; Zeng, Bin
- Abstract
The present study verified whether acyl-coenzyme A (acyl-CoA)-binding protein (ACBP) affected the production of <italic>Monascus</italic> pigments (MPs) in <italic>Monascus ruber</italic> CICC41233 (MrACBP). Phylogenetic analysis revealed that the cloned <italic>Mracbp</italic> gene, which encoded the MrACBP protein, exhibited the closest match (99% confidence level) to the gene from <italic>Penicilliopsis zonata</italic>. The MrACBP and maltose-binding protein (MBP) were simultaneously expressed in <italic>Escherichia coli</italic> 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 <italic>Mracbp</italic> gene was homologously overexpressed in <italic>M. ruber</italic> CICC41233, and a positive transformant <italic>M. ruber</italic> ACBP5 was isolated. The fatty acid myristic acid in <italic>M. ruber</italic> ACBP5 was lower than that in the parent strain <italic>M. ruber</italic> CICC41233. However, when compared with the parent strain, the production of total MPs, water-soluble pigment, and ethanol-soluble pigment in <italic>M. ruber</italic> 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 <italic>acbp</italic>, <italic>pks</italic>, <italic>mppr1</italic>, <italic>fasA</italic>, and <italic>fasB</italic> 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.
- Subjects
CARRIER proteins; PROTEIN binding; MONASCUS; PROTEIN expression; BACTERIAL proteins
- Publication
3 Biotech, 2018, Vol 8, Issue 2, p0
- ISSN
2190-572X
- Publication type
Article
- DOI
10.1007/s13205-018-1147-9