Directed evolution of an aminoalcohol dehydrogenase for efficient production of double chiral aminoalcohols

Urano, Nobuyuki; Fukui, Satoko; Kumashiro, Shoko; Ishige, Takeru; Kita, Shinji; Sakamoto, Keiji; Kataoka, Michihiko; Shimizu, Sakayu

doi:10.1016/j.jbiosc.2010.11.005

Cited by 20 publications

(15 citation statements)

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“…In particular, they produce NADP + -dependent aminoalcohol dehydrogenases (AADHs), including l-1-amino-2-propanol dehydrogenase (EC 1.1.1.75), which are also found in several other microorganisms [6,7]. The AADH of Rhodococcus erythropolis MAK154 exhibits unique stereospecificity and produces double chiral aminoalcohols [8][9][10], which are used as pharmaceuticals. Interestingly, the whole genomic sequence of R. erythropolis PR4 has been determined (GenBank accession no.…”

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confidence: 99%

Construction of Rhodococcus expression vectors and expression of the aminoalcohol dehydrogenase gene in Rhodococcus erythropolis

Yamamura

2018

Bioscience, Biotechnology, and Biochemistry

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NADP-dependent aminoalcohol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 produces double chiral aminoalcohols, which are used as pharmaceuticals. However, the genetic manipulation of Rhodococcus strains to increase their production of such industrially important enzymes is not well studied. Therefore, I aimed to construct Rhodococcus expression vectors, derived from the Rhodococcus-Escherichia coli shuttle vector pRET1102, to express aadh. The plasmid pRET1102 could be transformed into many actinomycete strains, including R. erythropolis. The transformation efﬁciency for a species closely related to R. erythropolis was higher than that for other actinomycete strains. Promoters of various strengths, hsp, 1200rep, and TRR, were obtained from Gram-positive bacteria. The activity of TRR was stronger than that of hsp and 1200rep. The aadh-expressing plasmid pRET1172 with TRR could be transformed into many actinomycete strains to increase their AADH production. The Rhodococcus expression vector, pRET11100, constructed by removing aadh from the pRET1172 plasmid may be useful for bioconversion.

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confidence: 99%

Construction of Rhodococcus expression vectors and expression of the aminoalcohol dehydrogenase gene in Rhodococcus erythropolis

Yamamura

2018

Bioscience, Biotechnology, and Biochemistry

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“…Although many SDRs have high enantioselectivity [19][20][21][22][23], CLA-DH had low enantioselectivity to dehydrogenate both (R) and (S) hydroxy fatty acids (Table1) and produce (R) and (S) hydroxy fatty acids from oxo fatty acid. In addition, CLA-DH dehydrogenated 10-, 12-, and 13-hydroxy fatty acids (Table 1), indicating its low regioselectivity.…”

Section: Discussionmentioning

confidence: 99%

Characterization of hydroxy fatty acid dehydrogenase involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a

Takeuchi

Kishino

Park

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

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Hydroxy fatty acid dehydrogenase, which is involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a, was cloned, expressed, purified, and characterized. The enzyme preferentially catalyzed NADH-dependent hydrogenation of oxo fatty acids over NAD +-dependent dehydrogenation of hydroxy fatty acids. In the dehydrogenation reaction, fatty acids with an internal hydroxy group such as 10-hydroxy-cis-12-octadecenoic acid, 12-hydroxy-cis-9-octadecenoic acid, and 13-hydroxy-cis-9-octadecenoic acid served as better substrates than those with α-or β-hydroxy groups such as 3-hydroxyoctadecanoic acid or 2-hydroxyeicosanoic acid. The apparent Km value for 10-hydroxy-cis-12-octadecenoic acid (HYA) was estimated to be 38 μM with a kcat of 7.6•10 −3 s −1. The apparent Km value for 10-oxo-cis-12-octadecenoic acid (KetoA) was estimated to be 1.8 μM with a kcat of 5.7•10 −1 s −1. In the hydrogenation reaction of KetoA, both (R)-and (S)-HYA were generated, indicating that the enzyme has low stereoselectivity. This is the first report of a dehydrogenase with a preference for fatty acids with an internal hydroxy group.

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“…In order to reduce the product inhibition of aminoalcohol dehydrogenase (AADH) from Rhodococcus erythropolis MAK154, Urano and coworkers screened a mutant library using a colorimetric assay in the presence of high concentrations of the product d -pseudoephedrine. As a result, a variant AADH carrying a double mutation was obtained, which showed higher activity in the presence of up to 100 mg/mL d -pseudoephedrine (Urano et al, 2011). In order to perform kinetic resolution of dl -propargylglycine, phenylalanine dehydrogenase was engineered by Chen and coworkers via directed evolution (Chen & Engel, 2009).…”

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confidence: 99%

Biocatalyst development by directed evolution

Wang

Zhao

2012

Bioresource Technology

121

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Biocatalysis has emerged as a great addition to traditional chemical processes for production of bulk chemicals and pharmaceuticals. To overcome the limitations of naturally occurring enzymes, directed evolution has become the most important tool for improving critical traits of biocatalysts such as thermostability, activity, selectivity, and tolerance towards organic solvents for industrial applications. Recent advances in mutant library creation and high-throughput screening have greatly facilitated the engineering of novel and improved biocatalysts. This review provides an update of the recent developments in the use of directed evolution to engineer biocatalysts for practical applications.

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Directed evolution of an aminoalcohol dehydrogenase for efficient production of double chiral aminoalcohols

Cited by 20 publications

References 24 publications

Construction of Rhodococcus expression vectors and expression of the aminoalcohol dehydrogenase gene in Rhodococcus erythropolis

Construction of Rhodococcus expression vectors and expression of the aminoalcohol dehydrogenase gene in Rhodococcus erythropolis

Characterization of hydroxy fatty acid dehydrogenase involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a

Biocatalyst development by directed evolution

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