Biochemical Characterization and Structural Analysis of a Bifunctional Cellulase/Xylanase from Clostridium thermocellum

Yuan, Shuo‐Fu; Wu, Tzu-Hui; Lee, Hsiao-Lin; Hsieh, Han-Yu; Lin, Wen-Ling; Yang, Barbara; Chang, Chien-Chih; Li, Qian; Gao, Jian; Huang, Chun‐Hsiang; Ho, Meng-Chiao; Guo, Rey‐Ting; Liang, Po‐Huang

doi:10.1074/jbc.m114.604454

Cited by 59 publications

(36 citation statements)

References 31 publications

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“…Apparently, when comparing the kinetic ratios (that is, V max /K m ) of cellulases with those of xylanases within a single organism, it emerged from our work that the ratios of cellulases were always higher than those of xylanases for all the obtained fungal isolates as described in Figure 5 and Table 3. This scenario is not unusual because previously, a recombinant protein from Clostridium thermocellum, CtCel5E, that had a dual function as a cellulase and xylanase, displayed a K m value of 2.1 mM and a V max of 1 564 µmol/s for the cellulase and a K m value of 4.6 mM and a V max of 883.5 µmol/s for the xylanase (Yuan et al, 2015). Notably, all the kinetic values of CtCel5E together with most of those proteins in Tables 5 and 6 were generally lower than those of our own enzyme extracts probably due to three possible reasons.…”

Section: Most Studies Nowadays Have Focused On Cellulases Andmentioning

confidence: 90%

Diversity of cellulase- and xylanase-producing filamentous fungi from termite mounds

Sibanda¹,

Ruzvidzo²,

Ncube³

et al. 2019

J. Yeast Fungal Res.

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Cellulases and xylanases are enzymes of industrial significance, particularly in the pulp, paper, textile, and animal feed industries. Moreover, their utilization in the food industry, among them, bakery, brewery, winery and fruit and vegetable juice production, cannot be underestimated. One of the potential sources of enzymes is the filamentous fungi, and hence bio-prospecting of this specific group of microorganisms with the highest levels of cellulase and xylanase secretions is being continuously undertaken. The specific aim of this study was to isolate and characterize cellulase-and xylanaseproducing filamentous fungi from termite mounds. Termite mounds have long been established as very good sources of filamentous fungi with the ability to secrete high levels of lignocellulolytic enzymes, and hence an ideal target for the bio-prospecting of cellulases and xylanases. In this study, various groups of filamentous fungi were isolated through enrichment and repeated sub-culturing. This was followed by screening using the Congo red plate-based assay. Cellulase and xylanase activities during the solid-state fermentation of wheat bran were detected and analyzed through spectrophotometry via the 3,5-dinitrosalicylic acid detection system for reducing sugars. The obtained fungal isolates were then finally characterized through zymography, reaction kinetics and morphological studies. Overall, a total of eight different groups of fungi, capable of decomposing cellulose and hemicellulose, were isolated, and their tentative identities established as Fusarium, Didymostible, Penicillium, Phytophthora, Oedocephalum, Aspergillus, Monosporascus and Acremonium. Taken together, findings of this study conceivably showed that termite mounds are a good source of filamentous fungi that in turn are also a good source of cellulases and xylanases that arguably, can be recommended for use in industrial and commercial settings.

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Section: Most Studies Nowadays Have Focused On Cellulases Andmentioning

confidence: 90%

Diversity of cellulase- and xylanase-producing filamentous fungi from termite mounds

Sibanda¹,

Ruzvidzo²,

Ncube³

et al. 2019

J. Yeast Fungal Res.

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“…Park et al (2014) reported 23-fold glucose production enhancement over that of free enzymes after their investigation of the effects of localization, surface accessibility, and functionality of synergetic enzymes on Scaf3-decorated bacteria outer membrane vesicles (OMVs) using phosphoric acid-swollen cellulose (PASC) as substrate. Yuan et al (2015), in an investigation to biochemically characterize and structurally analyze cellulase/xylanase from Clostridium thermocellum, also revealed equally insightful results. Advancement in cellulosome investigation has led to the advent of its artificial counterpart, called designer cellulosomes, described below.…”

Section: Cell-surface Display Systemsmentioning

confidence: 96%

“…On account of the obvious benefits reported in the literature, scientists have consistently investigated the gene (Yagüe et al 1990;Zverlov et al 2003;Koeck et al 2013), fusion/modification of enzymes (Ciolacu et al 2010;Lee et al 2010;Ye et al 2010;Lee et al 2011;Nakashima et al 2014), and the optimal growth (Islam et al 2013;Reed et al 2014) of these useful microbes to harness their inherent benefits. For example, the biochemical and biophysical characteristics of multimodular enzymes from Clostridium thermocellum (Zverlov et al 2005;Tachaapaikoon et al 2012;Brunecky et al 2012;Hirano et al 2013;Yuan et al 2015) and Thermotoga maritima (Chhabra et al 2002;Carvalho et al 2004;Pereira et al 2010;Wu et al 2011) have been reported.…”

Section: Bacteria Sourcesmentioning

confidence: 99%

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

Obeng

Adam

Budiman

et al. 2017

Bioresour. Bioprocess.

126

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The highly acclaimed prospect of renewable lignocellulosic biocommodities as obvious replacement of their fossilbased counterparts is burgeoning within the last few years. However, the use of the abundant lignocellulosic biomass provided by nature to produce value-added products, especially bioethanol, still faces significant challenges. One of the crucial challenging factors is in association with the expression levels, stability, and cost-effectiveness of the cellulose-degrading enzymes (cellulases). Interestingly, several recommendable endeavors in the bid to curb these challenges are in pursuance. However, the existing body of literature has not well provided the updated roadmap of the advancement and key players spearheading the current success. Moreover, the description of enzyme systems and emerging paradigms with high prospects, for example, the cell-surface display system has been ill-captured in the literature. This review focuses on the lignocellulosic biocommodity pathway, with emphasis on cellulase and hemicellulase systems. The paradigm shift towards cell-surface display system and its emerging recommendable developments have also been discussed. The attempts in supplementing cellulase with other enzymes, accessory proteins, and chemical additives have also been discussed. Moreover, some of the prominent and influential discoveries in the cellulase fraternity have been discussed.

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“…6,7 Progress in this field has seen the application of cell-free (crude extract), [8][9][10] immobilized [11][12][13] and microbial surface-displayed cellulases. [14][15][16] The cell-free form is the most common among the three categories. Recently, there has been a gradual shift towards the cell surface-display technology owing to its relatively cheaper production cost and reusability potential.…”

Section: Introductionmentioning

confidence: 99%

An optimal blend of single autodisplayed cellulases for cellulose saccharification – a proof of concept

Obeng

Ongkudon

Budiman

et al. 2018

J of Chemical Tech & Biotech

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BACKGOUND To date, it has been challenging to determine the optimal proportions of the individual cellulases in crude extracts and multienzyme systems to promote enzyme synergism. The application of single surface‐display of enzymes may obviate such challenges. This article presents a proof of concept of the optimal mixing of single surface‐displayed cellulases for cellulose hydrolysis using Simplex lattice mixture design. RESULTS The recently discovered maximized autotransporter‐mediated expression system was used to express the three cellulases. The biochemical screening analysis revealed that the enzymes have a broad range of functional pH (4–9) and temperature (30–100 °C) characteristics. The evaluated optima were pH 6 and 60 °C. Through the statistical design of experiment, a blend ratio of 1: 1.6: 1.4 of endoglucanase: exoglucanase: β‐glucosidase was identified as the ‘sweet spot’ for optimum sugar yield. The application of this blend generated about 0.354 mg mL‐1 and 0.446 mg mL‐1 of sugars from filter paper (Ø 6 mm, ∼2.5 mg) and 2.5% pretreated EFB, respectively, within 12 h. Supplementing the enzyme blend with CaCl2 and Tween 20 improved the sugar yield. CONCLUSION This research has revealed an interesting concept of efficiently addressing cellulase synergism. The overall outcome of this research is promising for the derivation of value from lignocellulose. © 2018 Society of Chemical Industry

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Biochemical Characterization and Structural Analysis of a Bifunctional Cellulase/Xylanase from Clostridium thermocellum

Cited by 59 publications

References 31 publications

Diversity of cellulase- and xylanase-producing filamentous fungi from termite mounds

Diversity of cellulase- and xylanase-producing filamentous fungi from termite mounds

Lignocellulases: a review of emerging and developing enzymes, systems, and practices

An optimal blend of single autodisplayed cellulases for cellulose saccharification – a proof of concept

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