Enzymatic synthesis of S-phenyl-l-cysteine from keratin hydrolysis industries wastewater with tryptophan synthase

Xu, Lisheng; Wang, Zhiyuan; Mao, Pingting; Liu, Junzhong; Zhang, Hongjuan; Liu, Qian; Jiao, Qingcai

doi:10.1016/j.biortech.2013.02.011

Cited by 11 publications

(5 citation statements)

References 9 publications

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“…However, the enzyme activity of P45A decreased significantly after 10 min in a solution of isopropanol, methanol, and glycol ( Figure 6B ), which was greater than that of GDH ( Figure 6A ). It may be that the hydrated layer and salt bridges of P45A were more easily destroyed than that of GDH, these are the major forces maintaining the structure of the protein ( Xu et al, 2013 ). On the other hand, the active pocket of P45A was more easily attacked by polar organic solvents as its higher hydrophobicity led to a decrease in the substrate binding ability ( Dumorne et al, 2017 ).…”

Section: Resultsmentioning

confidence: 99%

Directed evolution engineering to improve activity of glucose dehydrogenase by increasing pocket hydrophobicity

Qianqian²,

Yinbiao³

et al. 2022

Front. Microbiol.

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Glucose dehydrogenase (GDH) is a NAD(P)+ dependent oxidoreductase, which is useful in glucose determination kits, glucose biosensors, cofactor regeneration, and biofuel cells. However, the low efficiency of the catalysis hinders the use of GDH in industrial applications. In this study, an analysis of interactions between eight GDH mutants and NADP+ is powered by AlphaFold2 and Discovery Studio 3.0. The docking results showed that more hydrogen bonds formed between mutants, such as P45A and NADP+, which indicated that these mutants had the potential for high catalytic efficiency. Subsequently, we verified all the mutants by site-directed mutagenesis. It was notable that the enzyme activity of mutant P45A was 1829 U/mg, an improvement of 28-fold compared to wild-type GDH. We predicted the hydrophobicity of the protein-ligand complexes, which was confirmed by an 8-anilino-1-naphthalenesulphonic acid fluorescent probe. The following order of increasing hydrophobicity index was deduced: GDH < N46E < F155Y < P45A, which suggested that the enzyme activity of GDH is positively related to its pocket hydrophobicity. Furthermore, P45A still showed better catalytic ability in organic solvents, reaching 692 U/mg in 10% isopropanol, which was 19-fold that of the wild-type GDH. However, its substrate affinity was affected by organic solvents. This study provides a good theoretical foundation for further improving the catalytic efficiency of GDH.

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Section: Resultsmentioning

confidence: 99%

Directed evolution engineering to improve activity of glucose dehydrogenase by increasing pocket hydrophobicity

Qianqian²,

Yinbiao³

et al. 2022

Front. Microbiol.

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“…A series of analogues of L-tryptophan were synthesized through a β-reaction using tryptophan synthase [13]. Likewise, we reported the synthesis of S -phenyl-L-cysteine using tryptophan synthase from L-serine derived from the hydrolysis of keratin from industrial wastewater and thiophenol [14]. High-purity tryptophan was produced in excellent yield.…”

Section: Discussionmentioning

confidence: 99%

Highly efficient preparation of active S-phenyl-L-cysteine with tryptophan synthase using a chemoenzymatic method

Zhang

Gao

et al. 2019

BMC Biotechnol

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Background S-Phenyl-L-cysteine is regarded as having potential applicability as an antiretroviral/protease inhibitor for human immunodeficiency virus (HIV). In the present study, optically active S -phenyl-L-cysteine was prepared in a highly efficient manner from inexpensive bromobenzene using tryptophan synthase through a chemoenzymatic method. Results The chemoenzymatic method used a four-step reaction sequence. The process started with the reaction of magnesium and bromobenzene, followed by a Grignard reaction, and then hydrolysis and enzymatic synthesis using tryptophan synthase. Through this approach, S -phenyl-L-cysteine was chemoenzymatically synthesized using tryptophan synthase from thiophenol and L-serine as the starting material. Conclusions High-purity, optically active S -phenyl-L-cysteine was efficiently and inexpensively obtained in a total yield of 81.3% (> 99.9% purity).

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“…Directed evolution can promote the enzymatic activity of lipase, xylanase, a-galactosidases, monoamine oxidase and nitrilase [14][15][16][17][18][19]. L-tryptophan and S-phenyl-L-cysteine were synthesized by tryptophan synthase of Escherichia coli k-12, and it was found that tryptophan synthase has the potential to synthesize L-tryptophan and its derivatives [20][21][22]. In this study, directed evolution of tryptophan synthase was performed by error-prone PCR.…”

Section: Introductionmentioning

confidence: 99%

Directed Evolution Improves the Enzymatic Synthesis of L-5-Hydroxytryptophan by an Engineered Tryptophan Synthase

Huo

et al. 2021

Appl Biochem Biotechnol

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L-5-Hydroxytryptophan is an important amino acid that is widely used in food and medicine. In this study, L-5-hydroxytryptophan was synthesized by a modi ed tryptophan synthase. A direct evolution strategy was applied to engineer tryptophan synthase from Escherichia coli to improve the e ciency of L-5hydroxytryptophan synthesis. Tryptophan synthase was modi ed by error-prone PCR. A high activity mutant enzyme (V231A/K382G) was obtained by a high-throughput screening method. The activity of mutant enzyme (V231A/K382G) is 3.79 times higher than that of its parent, and k cat /K m of the mutant enzyme (V231A/K382G) was 4.36 mM − 1 •s − 1 . The mutant enzyme (V231A/K382G) reaction conditions for the production of L-5-hydroxytryptophan were 100 mmol/L L-serine at pH 8.5 and 35°C for 15 h, reaching a yield of L-5-hydroxytryptophan of 86.7%. Directed evolution is an effective strategy to increase the activity of tryptophan synthase.

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Enzymatic synthesis of S-phenyl-l-cysteine from keratin hydrolysis industries wastewater with tryptophan synthase

Cited by 11 publications

References 9 publications

Directed evolution engineering to improve activity of glucose dehydrogenase by increasing pocket hydrophobicity

Directed evolution engineering to improve activity of glucose dehydrogenase by increasing pocket hydrophobicity

Highly efficient preparation of active S-phenyl-L-cysteine with tryptophan synthase using a chemoenzymatic method

Directed Evolution Improves the Enzymatic Synthesis of L-5-Hydroxytryptophan by an Engineered Tryptophan Synthase

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