title: Cell surface engineering of α-l-rhamnosidase for naringin hydrolysis.
abstract: An α-l-rhamnosidase gene (rhaL1) containing an open reading frame of 2046-bp encoding a 681-amino acid protein (RhaL1) was cloned from Alternaria sp. L1 for naringin hydrolysis on the cell surface of Saccharomyces cerevisiae EBY-100. RhaL1 anchored to the yeast cell surface showed maximum enzyme activity at pH 6.0-6.5 and 70°C and was stable at pH 2.5-12.0 below 60°C. When the yeast cells were employed to hydrolyze naringin in grapefruit juice, about 85% naringin was hydrolyzed at 60°C in 10min. The yeast cells were harvested and recycled for the next batch. The hydrolysis rate of the naringin was maintained at over 80% for 10 batches. These results demonstrate the stability of the RhaL1-expressing yeast cells and effective in hydrolysis of naringin in juice. Thus, the system could have promise for industrial bitterness reduction.
authors.affiliation: State Key Lab of Microbial Technology and National Glycoengineering Research Center, Shandong University, Jinan 250100, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Lili
authors.initials: L
authors.lastname: Lu
authors.forename: Min
authors.initials: M
authors.lastname: Xiao
mesh_terms: D000528:Alternaria / Q000201:enzymology*; D000595:Amino Acid Sequence; D001628:Beverages; D002462:Cell Membrane / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D032083:Citrus paradisi; D003001:Cloning, Molecular; D004795:Enzyme Stability; D044950:Flavanones / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005800:Genes, Fungal / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013696:Temperature
chemical_list: D044950:Flavanones; D006026:Glycoside Hydrolases; C022525:alpha-L-rhamnosidase; C005274:naringin