2017
DOI: 10.1016/j.jbiotec.2016.11.034
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One-pot synthesis of glutathione by a two-enzyme cascade using a thermophilic ATP regeneration system

Abstract: In vitro cascade catalysis using enzyme-based system is becoming a promising biomanufacturing platform for biofuels and biochemicals production. Glutathione is a pivotal non-protein thiol compound and widely applied in food and pharmaceutical industries. In this study, glutathione was synthesized by a bifunctional glutathione synthetase together with a thermophilic ATP regeneration system through a two-enzyme cascade in vitro. Four bifunctional glutathione synthetases from Streptococcus sanguinis, S. gordonii,… Show more

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Cited by 36 publications
(13 citation statements)
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“…The plasmid pET28a‐GshF containing gshF from S. sanguinis and S. thermophiles were constructed previously 13,14 . Protein structure model of GshF was obtained from SWISS‐MODEL using GshF (PDB, 3LN6), which was from S. agalactiae , as a template (identity 65%) 7 .…”
Section: Methodsmentioning
confidence: 99%
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“…The plasmid pET28a‐GshF containing gshF from S. sanguinis and S. thermophiles were constructed previously 13,14 . Protein structure model of GshF was obtained from SWISS‐MODEL using GshF (PDB, 3LN6), which was from S. agalactiae , as a template (identity 65%) 7 .…”
Section: Methodsmentioning
confidence: 99%
“…Previous studies had shown that GshFs, which were derived from Streptococcus sanguinis (GshFSS) and Streptococcus thermophiles (GshFST), had superior ability to synthesize GSH 13,14 . In addition, structure‐based protein engineering and site‐directed mutagenesis are also available methods that could further enhance the enzymatic properties.…”
Section: Introductionmentioning
confidence: 99%
“…Whole‐cell biocatalysts were frequently adopted for ATP regeneration from ADP and the thermostability of the selected proteins allowed the biotransformations to run at high temperatures, with or without pre‐treatment of the enzymatic preparation to suppress unwanted side‐reactions and increase membrane permeability [34,42,53–54] . In vitro cascades, hybrid in vivo/in vitro cascades comprising purified proteins or more rarely reactions with cell‐free extracts were also exemplified.…”
Section: Application Of Polyphosphate Kinasesmentioning
confidence: 99%
“…On the one hand, reaction intensification attempts revealed that increasing concentration of polyP can have inhibitory effects on the PPK employed, the ATP‐consuming enzyme or both [34,42,54,59] . PolyP might exert its inhibitory effect either directly, for example by blocking enzymes’ active site (as described in section “Classification and Substrate Preference”), [32,43] or indirectly, for instance chelating divalent cations necessary for activity [41,54,59–63] . On the other hand, it was shown that an excess polyP is needed to drive ATP regeneration [8,36,60,62–64] .…”
Section: Application Of Polyphosphate Kinasesmentioning
confidence: 99%
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