Screening and selection of wild strains for L-arabinose isomerase production

Manzo, Ricardo Martín; Simonetta, A. C.; Rubiolo, Amelia Catalina; Mammarella, E

doi:10.1590/s0104-66322013000400003

Cited by 11 publications

(18 citation statements)

References 33 publications

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“…A microbiological method for the selection of wild-type strains producing L-arabinose isomerase was developed based on both the ability of the cells to produce acids and their capacity to grow on L-arabinose (Manzo et al, 2013). By using 0.5% of L-arabinose as the main carbon source and selected media, three strains could be found to be able to ferment L-arabinose: Enterococcus faecium DBFIQ ID: E36, E. faecium DBFIQ ID: ETW4 and Pediococcus acidilactici ATCC ID: 8042.…”

Section: Introductionmentioning

confidence: 99%

Whole cell biocatalysts: essential workers from Nature to the industry

Carvalho

2016

Microbial Biotechnology

213

151

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SummaryMicroorganisms have been exposed to a myriad of substrates and environmental conditions throughout evolution resulting in countless metabolites and enzymatic activities. Although mankind have been using these properties for centuries, we have only recently learned to control their production, to develop new biocatalysts with high stability and productivity and to improve their yields under new operational conditions. However, microbial cells still provide the best known environment for enzymes, preventing conformational changes in the protein structure in non‐conventional medium and under harsh reaction conditions, while being able to efficiently regenerate necessary cofactors and to carry out cascades of reactions. Besides, a still unknown microbe is probably already producing a compound that will cure cancer, Alzeihmer's disease or kill the most resistant pathogen. In this review, the latest developments in screening desirable activities and improving production yields are discussed.

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Section: Introductionmentioning

confidence: 99%

Whole cell biocatalysts: essential workers from Nature to the industry

Carvalho

2016

Microbial Biotechnology

213

151

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“…For this reason, the production of d -tagatose from d -galactose using l -AI has been studied in recent years [ 6 , 11 , 12 , 13 ]. However, the enzymatic process still has a number of bottlenecks that need to be overcome, e.g., low protein concentration in the fermentation step using wild-type-producing strains [ 14 , 15 ], low productivity in the isomerization reaction, and reduced thermostability of the enzyme [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

Engineering the l-Arabinose Isomerase from Enterococcus Faecium for d-Tagatose Synthesis

Sousa

Manzo

Garcı́a³

et al. 2017

Molecules

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l-Arabinose isomerase (EC 5.3.1.4) (l-AI) from Enterococcus faecium DBFIQ E36 was overproduced in Escherichia coli by designing a codon-optimized synthetic araA gene. Using this optimized gene, two N- and C-terminal His-tagged-l-AI proteins were produced. The cloning of the two chimeric genes into regulated expression vectors resulted in the production of high amounts of recombinant N-His-l-AI and C-His-l-AI in soluble and active forms. Both His-tagged enzymes were purified in a single step through metal-affinity chromatography and showed different kinetic and structural characteristics. Analytical ultracentrifugation revealed that C-His-l-AI was preferentially hexameric in solution, whereas N-His-l-AI was mainly monomeric. The specific activity of the N-His-l-AI at acidic pH was higher than that of C-His-l-AI and showed a maximum bioconversion yield of 26% at 50 °C for d-tagatose biosynthesis, with Km and Vmax parameters of 252 mM and 0.092 U mg−1, respectively. However, C-His-l-AI was more active and stable at alkaline pH than N-His-l-AI. N-His-l-AI follows a Michaelis-Menten kinetic, whereas C-His-l-AI fitted to a sigmoidal saturation curve.

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“…A positive impact on the cultivation and screening of enzymes can be obtained if the activity could be determined quickly and in real time (Manzo et al 2013). Other impurities, such as protein and carbohydrates, in a zymotic fluid can influence the accuracy of enzyme activity determination.…”

Section: Application Of the Modified Methods For Esterase Activity Detmentioning

confidence: 99%

Accurately Determining Esterase Activity via the Isosbestic Point of p-Nitrophenol

Peng¹,

Fu²,

Líu³

et al. 2016

BioResources

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Esterase is an important enzyme for ester hydrolysis or synthesis. Its activity, however, has not been accurately ascertained due to a lack of accurate protocols. In this study, the isosbestic point of p-nitrophenol was found and used as the marker for its activity. The methodology avoided decomposition of the substrate, chromophore agents, and pH changes. The esterase activity was determined accurately and rapidly in a complex solution. In this protocol system, organic solvents were used for dissolving substrates, which influenced activity determination to some extent. Among the solvents tested, methanol exerted the least inhibitory influence. The results indicated that this modified method has potential to be applied for esterase activity determination on a large scale and in real time.

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Screening and selection of wild strains for L-arabinose isomerase production

Cited by 11 publications

References 33 publications

Whole cell biocatalysts: essential workers from Nature to the industry

Whole cell biocatalysts: essential workers from Nature to the industry

Engineering the l-Arabinose Isomerase from Enterococcus Faecium for d-Tagatose Synthesis

Accurately Determining Esterase Activity via the Isosbestic Point of p-Nitrophenol

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