Direct screening of libraries of yeast clones for alpha-amylase activity on raw starch hydrolysis.
High-throughput screening for high-activity barley alpha-amylase mutants expressed in Saccharomyces cerevisiae is hampered by the interference of reducing agents, particularly the glucose used in yeast growth media. The present investigation employed colorimetric and chemiluminescent detection systems that enable direct and rapid screening of activities on raw starch substrate. Active clones could be separated into two groups, based on high total activity or high specific activity.
Wong
Dominic W S
DW
Western Regional Research Center, USDA-Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710, USA. dwsw@pw.usda.gov
Batt
Sarah B
SB
Lee
Charles C
CC
Robertson
George H
GH
D001483:Base Sequence; D003001:Cloning, Molecular; D004789:Enzyme Activation; D015723:Gene Library; D005820:Genetic Testing; D005990:Glycerol; D006868:Hydrolysis; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009154:Mutation; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D013213:Starch / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D013213:Starch; D000516:alpha-Amylases; D005990:Glycerol