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Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

ABSTRACT: Tannase (tannin acyl hydrolase E.C 3.1.1.20) is an inducible, largely extracellular enzyme that causes the hydrolysis of ester and depside bonds present in various substrates. Large scale industrial application of this enzyme is very limited owing to its high production costs. In the present study, cost effective production of tannase by Klebsiella pneumoniae KP715242 was studied under submerged fermentation using different tannin rich agro-residues like Indian gooseberry leaves (Phyllanthus emblica), Black plum leaves (Syzygium cumini), Eucalyptus leaves (Eucalyptus glogus) and Babul leaves (Acacia nilotica). Among all agro-residues, Indian gooseberry leaves were found to be the best substrate for tannase production under submerged fermentation. Sequential optimization approach using Taguchi orthogonal array screening and response surface methodology was adopted to optimize the fermentation variables in order to enhance the enzyme production. Eleven medium components were screened primarily by Taguchi orthogonal array design to identify the most contributing factors towards the enzyme production. The four most significant contributing variables affecting tannase production were found to be pH (23.62 %), tannin extract (20.70 %), temperature (20.33 %) and incubation time (14.99 %). These factors were further optimized with central composite design using response surface methodology. Maximum tannase production was observed at 5.52 pH, 39.72 °C temperature, 91.82 h of incubation time and 2.17 % tannin content. The enzyme activity was enhanced by 1.26 fold under these optimized conditions. The present study emphasizes the use of agro-residues as a potential substrate with an aim to lower down the input costs for tannase production so that the enzyme could be used proficiently for commercial purposes.

SUBMITTER: Kumar M

PROVIDER: S-EPMC4943918 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

Kumar Mukesh M Singh Amrinder A Beniwal Vikas V Salar Raj Kumar RK

AMB Express 20160713 1

Tannase (tannin acyl hydrolase E.C 3.1.1.20) is an inducible, largely extracellular enzyme that causes the hydrolysis of ester and depside bonds present in various substrates. Large scale industrial application of this enzyme is very limited owing to its high production costs. In the present study, cost effective production of tannase by Klebsiella pneumoniae KP715242 was studied under submerged fermentation using different tannin rich agro-residues like Indian gooseberry leaves (Phyllanthus emb ...[more]

PMID: 27411334

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Project description:ObjectiveMulberry (Morus spp.) fruits and leaves have been proven to possess nutraceutical properties. Due to its fast and easy growing characteristics, mulberry fruits (MF) and leaves (ML) potentially emerge as a great source of functional foods. This study aims to enhance bioactivities (antioxidant, anti-inflammation, and hypoglycemic activity) of MF and ML via submerged fermentation using bacteria (Lactobacillus plantarum TAR 4), yeast (Baker's yeast and red yeast) and fungi (Tempeh and Tapai starter).MethodsIn this study, 25% (mass to volume ratio) of MF and ML were fermented (48 h) with 1% (mass to volume ratio) of different microbial cultures, respectively. Effects of different fermentations on MF and ML were determined based on the changes of total phenolics (TPC), flavonoids (TFC), anthocyanins, total sugar, DPPH activity, ferric reducing antioxidant power (FRAP), albumin denaturation inhibition activity (ADI), anti-lipoxygenase activity and α-amylase inhibition activity (AI).ResultsGenerally, ML had higher AI than MF. However, MF exhibited higher DPPH, FRAP and anti-lipoxygenase activity than ML. After all forms of fermentation, DPPH and AI activity of MF and ML were increased significantly (P < 0.05). However, the effects of fermentation on TPC, FRAP, ADI and anti-lipoxygenase activity of MF were in contrast with ML. TPC, FRAP and anti-lipoxygenase activity of ML were enhanced, but reduced in MF after fermentation. Although the effects exerted by different microorganisms in MF and ML fermentation were different, the bioactivities of MF and ML were generally improved after fermentation. Fermentation by Tempeh starter enhanced TPC (by 2-fold), FRAP (by 2.3-fold), AI (at 10% increment) and anti-lipoxygenase activity (by 5-fold) of ML, whereas Tapai fermentation effectively enhanced the DPPH (at 17% increment) and ADI (by 2-fold) activity of MF.ConclusionFindings of this study provide an insight into the future process design of MF and ML processing into novel functional foods.

Project description:BACKGROUND:It has long been recognized that Klebsiella pneumoniae can grow anaerobically on citrate. Genes responsible for citrate fermentation of K. pneumoniae were known to be located in a 13-kb gene cluster on the chromosome. By whole genome comparison of the available K. pneumoniae sequences (MGH 78578, 342, and NTUH-K2044), however, we discovered that the fermentation gene cluster was present in MGH 78578 and 342, but absent in NTUH-K2044. In the present study, the previously unknown genome diversity of citrate fermentation among K. pneumoniae clinical isolates was investigated. RESULTS:Using a genomic microarray containing probe sequences from multiple K. pneumoniae strains, we investigated genetic diversity among K. pneumoniae clinical isolates and found that a genomic region containing the citrate fermentation genes was not universally present in all strains. We confirmed by PCR analysis that the gene cluster was detectable in about half of the strains tested. To demonstrate the metabolic function of the genomic region, anaerobic growth of K. pneumoniae in artificial urine medium (AUM) was examined for ten strains with different clinical histories and genomic backgrounds, and the citrate fermentation potential was found correlated with the genomic region. PCR detection of the genomic region yielded high positive rates among a variety of clinical isolates collected from urine, blood, wound infection, and pneumonia. Conserved genetic organizations in the vicinity of the citrate fermentation gene clusters among K. pneumoniae, Salmonella enterica, and Escherichia coli suggest that the 13-kb genomic region were not independently acquired. CONCLUSION:Not all, but nearly half of the K. pneumoniae clinical isolates carry the genes responsible for anaerobic growth on citrate. Genomic variation of citrate fermentation genes in K. pneumoniae may contribute to metabolic diversity and adaptation to variable nutrient conditions in different environments.

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Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

Publications

Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

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