Identification and Application of Enantiocomplementary Lactamases for Vince Lactam Derivatives

Assaf, Zeinab; Eger, E. I.; Vitnik, Željko J.; Fabian, Walter M. F.; Ribitsch, Doris; Guebitz, Georg M.; Faber, Kurt; Hall, Mélanie

doi:10.1002/cctc.201402077

Cited by 22 publications

(14 citation statements)

References 36 publications

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“…A γ-lactamase mutant (designated as MhIHL-V54L hereafter) from Microbacterium hydrocarbonoxydans was selected as a model biocatalyst and placed in the interior of the nanocage. γ-Lactamase is a versatile enzyme that is widely used for the enzymatic resolution of racemic 2-azabicyclo [2.2.1] hept-5-en-3-one (Vince lactam) [ 17 – 21 ]. The chiral products are valuable synthons used for an increasing number of drug candidates, such as Ziagen (Abacavir) [ 19 ].…”

Section: Resultsmentioning

confidence: 99%

Construction of an organelle-like nanodevice via supramolecular self-assembly for robust biocatalysts

Zheng

Zhu

2018

Microb Cell Fact

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BackgroundWhen using the microbial cell factories for green manufacturing, several important issues need to be addressed such as how to maintain the stability of biocatalysts used in the bioprocess and how to improve the synthetic efficiency of the biological system. One strategy widely used during natural evolution is the creation of organelles which can be used for regional control. This kind of compartmentalization strategy has inspired the design of artificial organelle-like nanodevice for synthetic biology and “green chemistry”.ResultsMimicking the natural concept of functional compartments, here we show that the engineered thermostable ketohydroxyglutarate aldolase from Thermotoga maritima could be developed as a general platform for nanoreactor design via supramolecular self-assembly. An industrial biocatalyst-(+)-γ-lactamase was selected as a model catalyst and successful encapsulated in the nanoreactor with high copies. These nanomaterials could easily be synthesized by Escherichia coli by heterologous expression and subsequently self-assembles into the target organelle-like nanoreactors both in vivo and in vitro. By probing their structural characteristics via transmission electronic microscopy and their catalytic activity under diverse conditions, we proved that these nanoreactors could confer a significant benefit to the cargo proteins. The encapsulated protein exhibits significantly improved stability under conditions such as in the presence of organic solvent or proteases, and shows better substrate tolerance than free enzyme.ConclusionsOur biodesign strategy provides new methods to develop new catalytically active protein-nanoreactors and could easily be applied into other biocatalysts. These artificial organelles could have widely application in sustainable catalysis, synthetic biology and could significantly improve the performance of microbial cell factories.Graphical Abstract Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0873-3) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Construction of an organelle-like nanodevice via supramolecular self-assembly for robust biocatalysts

Zheng

Zhu

2018

Microb Cell Fact

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“…Over the past few decades, intensive efforts have been made to discover effective γ-lactamases by both screening and genome mining methods. To date, several (+)-γ-lactamases and (−)-γ-lactamases have been reported 6 7 8 9 10 11 12 13 14 15 . Although the three currently known (−)-γ-lactamases (1HKH_A from an Aureobacterium species, ACY56506.1 from Microbacterium hydrocarbonoxydan s and WP_014491492.1 from Bradyrhizobium japonicum ) are similar in sequence to each other, the amino acid sequence are quite diverse among the known (+)-γ-lactamases due to the different subfamilies of (+)-γ-lactamases.…”

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confidence: 99%

Structural insights into the γ-lactamase activity and substrate enantioselectivity of an isochorismatase-like hydrolase from Microbacterium hydrocarbonoxydans

Gao

Zhou

Zhang

et al. 2017

Sci Rep

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(+)-γ-lactamase catalyzes the specific hydrolysis of (+)-γ-lactam out of the racemic γ-lactam (2-Azabicyclo[2.2.1]hept-5-en-3-one) to leave optically pure (−)-γ-lactam, which is the key building block of antiviral drugs such as carbovir and abacavir. However, no structural data has been reported on how the enzymes bind the γ-lactams and achieve their enantioselectivities. We previously identified an isochorismatase-like hydrolase (IHL, Mh33H4-5540) with (+)-γ-lactamase activity, which constitutes a novel family of γ-lactamase. Here, we first discovered that this enzyme actually hydrolyzed both (+)- and (−)-γ-lactam, but with apparently different specificities. We determined the crystal structures of the apo-form, (+)-γ-lactam bound, and (−)-γ-lactam bound forms of the enzyme. The structures showed that the binding sites of both (+) and (−)-γ-lactam resemble those of IHLs, but the “cover” loop conserved in IHLs is lacking in the enzyme, probably resulting in its incomplete enantioselectivity. Structural, biochemical, and molecular dynamics simulation studies demonstrated that the steric clash caused by the binding-site residues, especially the side-chain of Cys111 would reduce the binding affinity of (−)-γ-lactam and possibly the catalytic efficiency, which might explain the different catalytic specificities of the enantiomers of γ-lactam. Our results would facilitate the directed evolution and application of Mh33H4-5540 in antiviral drug synthesis.

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“…Over the past 30 years, numerous lactamases have been isolated and applied in the ring-opening of Vince-lactam. [103][104][105][106][107][108][109][110][111][112][113][114][115][116][117][118] Resolution could, alternatively, be achieved using a variety of other enzymes including CAL-B and a non-haem chloroperoxidase from Streptomyces viridochromogenes. [119][120][121][122] CAL-B resolution efficiency could be increased by making use of the activated N-hydroxymethyl Vince-lactam.…”

Section: Scheme 10mentioning

confidence: 99%

Biocatalytic routes to anti-viral agents and their synthetic intermediates

Slagman

Fessner

2021

Chem. Soc. Rev.

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Identification and Application of Enantiocomplementary Lactamases for Vince Lactam Derivatives

Cited by 22 publications

References 36 publications

Construction of an organelle-like nanodevice via supramolecular self-assembly for robust biocatalysts

Construction of an organelle-like nanodevice via supramolecular self-assembly for robust biocatalysts

Structural insights into the γ-lactamase activity and substrate enantioselectivity of an isochorismatase-like hydrolase from Microbacterium hydrocarbonoxydans

Biocatalytic routes to anti-viral agents and their synthetic intermediates

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