RRC ID 48801
著者 Wang M, Deng WW, Zhang ZZ, Yu O.
タイトル Engineering an ABC Transporter for Enhancing Resistance to Caffeine in Saccharomyces cerevisiae.
ジャーナル J Agric Food Chem
Abstract In addressing caffeine toxicity to the producing cells, engineering a transporter that can move caffeine from cytoplasm across the cell membrane to the extracellular space, thus enhancing caffeine resistance and potentially increasing the yield in yeast, is important. An ABC-transporter bfr1 from Schizosaccharomyces pombe was cloned and transformed into S. cerevisiae, resulting in enhancing caffeine resistance. Afterward, a library of randomly mutagenized bfr1 mutants through error-prone PCR was generated. One mutant was identified with drastically increased caffeine resistance (15 mg/mL). Sequencing and structural analysis illustrated that many of the mutations occurred at the cytosolic domain. Site-directed mutagenesis of these mutations confirmed at least one amino acid that conferred enhancing caffeine resistance in the mutated bfr1. These data demonstrated engineering ABC-transporters can be an efficient way to reduce product toxicity in heterologous systems.
巻・号 64(42)
ページ 7973-7978
公開日 2016-10-26
DOI 10.1021/acs.jafc.6b03980
PMID 27696877
IF 4.192
引用数 1
リソース情報
遺伝子材料 S. pombe ORFeome clone SpYFH49H06 (SPW099774)