Work

Combinational biosynthesis and characterization of a fluorescent 82β-phycocyanin of Spirulina platensis

Year: 2012

Type: article

Abstract: To biosynthesize fluorescent Spirulina platensis ( Sp ) β-phycocyanin (PC) in Escherichia coli , a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 ( S6 ) proteome. A highly homologous gene, slr2049 , was obtained from the S6 genome. Sites 82 and 153 in β-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E . coli BL21: (i) pCDF- cpcB (C153A)- slr2049 - sll0583 - ho1 - pcyA ; and (ii) pCDF- cpcB (C82I)- slr2049 - sll0583 - ho1 - pcyA . Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82β-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology.

Source: Chinese Science Bulletin

DOI: 10.1007/s11434-012-5264-2

Authors: XiangYu Guan, WeiJie Zhang, XiaoYuan Chi, HanZhi Lin, JinFeng Wang, Song Qin