Characterization of Four New Distinct ω-Transaminases from Pseudomonas putida NBRC 14164 for Kinetic Resolution of Racemic Amines and Amino Alcohols

Wu, Hua-Lei; Zhang, Jiandong; Zhang, Chao-Feng; Fan, Xiaojun; Chang, Hong; Wei, Wuran

doi:10.1007/s12010-016-2263-9

Cited by 28 publications

(18 citation statements)

References 40 publications

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“…While a number of other discovery approaches have emerged in recent years, such as culture-independent metagenome screening, novel enzymes continue to be discovered in this way ( Fig. 1) (Wilding et al 2015(Wilding et al , 2016Wu et al 2017;Noshahri et al 2019). In the latter example by Noshahri et al, the decades-old approach of using enrichment media with 1-PEA as the sole nitrogen source was given a new lease of life by applying this method to microorganisms from challenging environments.…”

Section: Culture-based Enzyme Discoverymentioning

confidence: 99%

Transaminases for industrial biocatalysis: novel enzyme discovery

Kelly

Mix

Moody

et al. 2020

Appl Microbiol Biotechnol

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Transaminases (TAms) are important enzymes for the production of chiral amines for the pharmaceutical and fine chemical industries. Novel TAms for use in these industries have been discovered using a range of approaches, including activity-guided methods and homologous sequence searches from cultured microorganisms to searches using key motifs and metagenomic mining of environmental DNA libraries. This mini-review focuses on the methods used for TAm discovery over the past two decades, analyzing the changing trends in the field and highlighting the advantages and drawbacks of the respective approaches used. This review will also discuss the role of protein engineering in the development of novel TAms and explore possible directions for future TAm discovery for application in industrial biocatalysis. Key Points• The past two decades of TAm enzyme discovery approaches are explored.• TAm sequences are phylogenetically analyzed and compared to other discovery methods.• Benefits and drawbacks of discovery approaches for novel biocatalysts are discussed.• The role of protein engineering and future discovery directions is highlighted.

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Section: Culture-based Enzyme Discoverymentioning

confidence: 99%

Transaminases for industrial biocatalysis: novel enzyme discovery

Kelly

Mix

Moody

et al. 2020

Appl Microbiol Biotechnol

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“…In contrast to Fold type I ( S )‐selective ω‐TAs, Fold type IV ( R )‐selective ω‐TAs have no reported activity toward β‐ or γ‐amino acids . Enzymes from both superfamilies show activity toward the small‐sized α‐alanine .…”

Section: Discussionmentioning

confidence: 99%

The ω‐transaminase engineering database (oTAED): A navigation tool in protein sequence and structure space

et al. 2018

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The ω-Transaminase Engineering Database (oTAED) was established as a publicly accessible resource on sequences and structures of the biotechnologically relevant ω-transaminases (ω-TAs) from Fold types I and IV. The oTAED integrates sequence and structure data, provides a classification based on fold type and sequence similarity, and applies a standard numbering scheme to identify equivalent positions in homologous proteins. The oTAED includes 67 210 proteins (114 655 sequences) which are divided into 169 homologous families based on global sequence similarity. The 44 and 39 highly conserved positions which were identified in Fold type I and IV, respectively, include the known catalytic residues and a large fraction of glycines and prolines in loop regions, which might have a role in protein folding and stability. However, for most of the conserved positions the function is still unknown. Literature information on positions that mediate substrate specificity and stereoselectivity was systematically examined. The standard numbering schemes revealed that many positions which have been described in different enzymes are structurally equivalent. For some positions, multiple functional roles have been suggested based on experimental data in different enzymes. The proposed standard numbering schemes for Fold type I and IV ω-TAs assist with analysis of literature data, facilitate annotation of ω-TAs, support prediction of promising mutation sites, and enable navigation in ω-TA sequence space. Thus, it is a useful tool for enzyme engineering and the selection of novel ω-TA candidates with desired biochemical properties.

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“…Resolution of racemic mixtures has been also achieved through the use of purified enzymes. Wu et al [24] demonstrated that ω-transaminase from P. putida NBRC 14164 entailed high enantioselectivity to different racemic amines and amino alcohols, including α-methylbenzylamine and 2-amino-2-phenylethanol. The enzyme PpspuC achieved ee > 99.0% of (R)-α-methylbenzylamine and (S)-2-amino-2-phenylethanol within 4 h and 24 h respectively, while the removal of racemic α-methylbenzylamine was 50.2% and that of racemic 2-amino-2-phenylethanol reached 50.7%.…”

Section: Biocatalytic Resolution Of Lupanine Enantiomeric Mixturementioning

confidence: 99%

Resolution of alkaloid racemate: a novel microbial approach for the production of enantiopure lupanine via industrial wastewater valorization

et al. 2020

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Background: Lupanine is a plant toxin contained in the wastewater of lupine bean processing industries, which could be used for semi-synthesis of various novel high added-value compounds. This paper introduces an environmental friendly process for microbial production of enantiopure lupanine.Results: Previously isolated P. putida LPK411, R. rhodochrous LPK211 and Rhodococcus sp. LPK311, holding the capacity to utilize lupanine as single carbon source, were employed as biocatalysts for resolution of racemic lupanine. All strains achieved high enantiomeric excess (ee) of L-(−)-lupanine (> 95%), while with the use of LPK411 53% of the initial racemate content was not removed. LPK411 fed with lupanine enantiomers as single substrates achieved 92% of D-(+)-lupanine biodegradation, whereas L-(−)-lupanine was not metabolized. Monitoring the transcriptional kinetics of the luh gene in cultures supplemented with the racemate as well as each of the enantiomers supported the enantioselectivity of LPK411 for D-(+)-lupanine biotransformation, while (trans)-6-oxooctahydro-1H-quinolizine-3-carboxylic acid was detected as final biodegradation product from D-(+)-lupanine use. Ecotoxicological assessment demonstrated that lupanine enantiomers were less toxic to A. fischeri compared to the racemate exhibiting synergistic interaction. Conclusions:The biological chiral separation process of lupanine presented here constitutes an eco-friendly and low-cost alternative to widely used chemical methods for chiral separation.

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Characterization of Four New Distinct ω-Transaminases from Pseudomonas putida NBRC 14164 for Kinetic Resolution of Racemic Amines and Amino Alcohols

Cited by 28 publications

References 40 publications

Transaminases for industrial biocatalysis: novel enzyme discovery

Transaminases for industrial biocatalysis: novel enzyme discovery

The ω‐transaminase engineering database (oTAED): A navigation tool in protein sequence and structure space

Resolution of alkaloid racemate: a novel microbial approach for the production of enantiopure lupanine via industrial wastewater valorization

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