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Literature summary for 1.14.99.63 extracted from

  • Zhou, P.; Li, M.; Shen, B.; Yao, Z.; Bian, Q.; Ye, L.; Yu, H.
    Directed coevolution of beta-carotene ketolase and hydroxylase and its application in temperature-regulated biosynthesis of astaxanthin (2019), J. Agric. Food Chem., 67, 1072-1080 .
    View publication on PubMed
Search for more literature for 1.14.99.63

Cloned(Commentary)

Cloned (Comment) Organism
gene ObktM, recombinant expression of wild-type and mutant enzymes in Saccharomaces cerevisiae strain BY4741, coexpression with gene OcrtZ encoding a beta-carotene 3-hydroxylase Haematococcus lacustris

Protein Variants

Protein Variants Comment Organism
H165R/V264D/F298Y/M1T/N188D/L271R site-directed mutagenesis, mutant OBKTM29 shows the best performance among all OBKTM mutants obtained by directed coevolution Haematococcus lacustris
additional information construction of an efficient astaxanthin-producing Saccharomyces cerevisiae strain BY4741 by combining protein engineering and dynamic metabolic regulation. First, superior mutants of beta-carotene ketolase and beta-carotene hydroxylase are obtained by directed coevolution to accelerate the conversion of beta-carotene to astaxanthin. Subsequently, the Gal4M9-based temperature-responsive regulation system is introduced to separate astaxanthin production from cell growth. Finally, 235 mg/l of (3S,3'S)-astaxanthin is produced by two-stage, high-density fermentation. Color-based high-throughput screening for directed coevolution of beta-carotene ketolase and beta-carotene hydroxylase Haematococcus lacustris

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
beta-carotene + 2 O2 + 2 reduced acceptor Haematococcus lacustris
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 echinenone + 3 H2O + 2 acceptor
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 ?
echinenone + 2 O2 + 2 reduced acceptor Haematococcus lacustris
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 canthaxanthin + 3 H2O + 2 acceptor
-
 ?
additional information Haematococcus lacustris poor efficiency of beta-carotene ketolation and hydroxylation, as well as the adverse effect of astaxanthin accumulation on cell growth, in vivo. Canthxanthin is produced by beta-carotene ketolase, ObktM, from beta-carotene, canthaxanthin is then further metabolized by a beta-carotene hydroxylase, CrtZ, in the engineered yeast strain expressing both enzymes from Haematococcus pluvialis and Xanthophyllomyces dendrorhous, respectively ?
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Organism

Organism UniProt Comment Textmining
Haematococcus lacustris
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 Haematococcus pluvialis
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-carotene + 2 O2 + 2 reduced acceptor
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 Haematococcus lacustris echinenone + 3 H2O + 2 acceptor
-
 ?
echinenone + 2 O2 + 2 reduced acceptor
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 Haematococcus lacustris canthaxanthin + 3 H2O + 2 acceptor
-
 ?
additional information poor efficiency of beta-carotene ketolation and hydroxylation, as well as the adverse effect of astaxanthin accumulation on cell growth, in vivo. Canthxanthin is produced by beta-carotene ketolase, ObktM, from beta-carotene, canthaxanthin is then further metabolized by a beta-carotene hydroxylase, CrtZ, in the engineered yeast strain expressing both enzymes from Haematococcus pluvialis and Xanthophyllomyces dendrorhous, respectively Haematococcus lacustris ?
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Synonyms

Synonyms Comment Organism
beta-carotene ketolase
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 Haematococcus lacustris
ObktM
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 Haematococcus lacustris
OBKTM29 mutant enzyme Haematococcus lacustris

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 in vivo assay at Haematococcus lacustris