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The identification and use of robust transaminases from a domestic drain metagenome.

ABSTRACT: Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identification, cloning and screening of 29 novel transaminases from a household drain metagenome. The most promising enzymes were fully characterised and the effects of pH, temperature, amine donor concentration and co-solvent determined. Several enzymes demonstrated good substrate tolerance as well as an unprecedented robustness for a wild-type transaminase. One enzyme in particular readily accepted IPA as an amine donor giving the same conversion with 2-50 equivalents, as well as being tolerant to a number of co-solvents, and operational in up to 50% DMSO - a characteristic as yet unobserved in a wild-type transaminase. This work highlights the value of using metagenomics for biocatalyst discovery from niche environments, and here has led to the identification of one of the most robust native transaminases described to date, with respect to IPA and DMSO tolerance.

SUBMITTER: Leipold L

PROVIDER: S-EPMC6394892 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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The identification and use of robust transaminases from a domestic drain metagenome.

Leipold Leona L Dobrijevic Dragana D Jeffries Jack W E JWE Bawn Maria M Moody Thomas S TS Ward John M JM Hailes Helen C HC

Green chemistry : an international journal and green chemistry resource : GC 20181115 1

Transaminases remain one of the most promising biocatalysts for use in chiral amine synthesis, however their industrial implementation has been hampered by their general instability towards, for example, high amine donor concentrations and organic solvent content. Herein we describe the identification, cloning and screening of 29 novel transaminases from a household drain metagenome. The most promising enzymes were fully characterised and the effects of pH, temperature, amine donor concentration ...[more]

PMID: 30930686

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Project description:BACKGROUND:Currently, many surgeons place a prophylactic drain in the abdominal or pelvic cavity after colorectal anastomosis as a conventional treatment. However, some trials have demonstrated that this procedure may not be beneficial to the patients. OBJECTIVE:To determine whether prophylactic placement of a drain in colorectal anastomosis can reduce postoperative complications. METHODS:We systematically searched all the electronic databases for randomized controlled trials (RCTs) that compared routine use of drainage to non-drainage regimes after colorectal anastomosis, using the terms "colorectal" or "colon/colonic" or "rectum/rectal" and "anastomo*" and "drain or drainage." Reference lists of relevant articles, conference proceedings, and ongoing trial databases were also screened. Primary outcome measures were clinical and radiological anastomotic leakage. Secondary outcome measures included mortality, wound infection, re-operation, and respiratory complications. We assessed the eligible studies for risk of bias using the Cochrane Risk of Bias Tool. Two authors independently extracted data. RESULTS:Eleven RCTs were included (1803 patients in total, 939 patients in the drain group and 864 patients in the no drain group). Meta-analysis showed that there was no statistically significant differences between the drain group and the no drain group in (1) overall anastomotic leakage (relative risk (RR)?=?1.14, 95 % confidence interval (CI) 0.80-1.62, P?=?0.47), (2) clinical anastomotic leakage (RR?=?1.39, 95 % CI 0.80-2.39, P?=?0.24), (3) radiologic anastomotic leakage (RR?=?0.92, 95 % CI 0.56-1.51, P?=?0.74), (4) mortality (RR?=?0.94, 95 % CI 0.57-1.55, P?=?0.81), (5) wound infection (RR?=?1.19, 95 % CI 0.84-1.69, P?=?0.34), (6) re-operation (RR?=?1.18, 95 % CI 0.75-1.85, P?=?0.47), and (7) respiratory complications (RR?=?0.82, 95 % CI 0.55-1.23, P?=?0.34). CONCLUSIONS:Routine use of prophylactic drainage in colorectal anastomosis does not benefit in decreasing postoperative complications.

Dataset Information

The identification and use of robust transaminases from a domestic drain metagenome.

Publications

The identification and use of robust transaminases from a domestic drain metagenome.

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