Expression of Rumen Microbial Fibrolytic Enzyme Genes in Probiotic
            <i>Lactobacillus reuteri</i>

Liu, Je-Ruei; Yu, Bin; Liu, Fu-Hwa; Cheng, K.-J.; Zhao, Xin

doi:10.1128/aem.71.11.6769-6775.2005

Cited by 43 publications

(48 citation statements)

References 39 publications

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“…intestinal probiotic strains (49)(50)(51). Recently, coexpression of a ␤-glucanase and a xylanase in Lactobacillus reuteri has been reported (51), and the transformed strain exhibited enzymatic activity on soluble ␤-glucan and xylan.…”

Section: Figmentioning

confidence: 99%

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Moraïs

Shterzer

Grinberg

et al. 2013

Appl Environ Microbiol

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Lactobacillus plantarum is an attractive candidate for bioprocessing of lignocellulosic biomass due to its high metabolic variability, including its ability to ferment both pentoses and hexoses, as well as its high acid tolerance, a quality often utilized in industrial processes. This bacterium grows naturally on biomass; however, it lacks the inherent ability to deconstruct lignocellulosic substrates. As a first step toward engineering lignocellulose-converting lactobacilli, we have introduced genes coding for a GH6 cellulase and a GH11 xylanase from a highly active cellulolytic bacterium into L. plantarum. For this purpose, we employed the recently developed pSIP vectors for efficient secretion of heterologous proteins. Both enzymes were secreted by L. plantarum at levels estimated at 0.33 nM and 3.3 nM, for the cellulase and xylanase, respectively, in culture at an optical density at 600 nm (OD 600 ) of 1. Transformed cells demonstrated the ability to degrade individually either cellulose or xylan and wheat straw. When mixed together to form a two-strain cell-based consortium secreting both cellulase and xylanase, they exhibited synergistic activity in the overall release of soluble sugar from wheat straw. This result paves the way toward metabolic harnessing of L. plantarum for novel biorefining applications, such as production of ethanol and polylactic acid directly from plant biomass.

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Section: Figmentioning

confidence: 99%

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Moraïs

Shterzer

Grinberg

et al. 2013

Appl Environ Microbiol

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“…Important preference for T in the third position was found in xynB from N. patriciarum and G was excluded from the wobble position . The reiterated sequences of RTTT determined in fungal xynR8 (Liu et al 2005) were also partially observed in the xylanase investigated here at the 197, 204 and 209 position of the deduced amino acid sequence (Fig. 3) with valin and arginine substitution in first two reiterated regions.…”

Section: Discussionmentioning

confidence: 82%

“…Rumen fungal enzymes have much interest for scientific investigation (Qiu et al 2000), because of the wide substrate specificity of these enzymes (Trinci et al 1994) and remarkably powerful fibrolytic activity (Liu et al 2005). Since the production of fibrolytic enzymes of anaerobic fungi is under the control of catabolite repression (Mountfort & Asher 1983), in order to stimulate the xylanase expression in our isolate Neocallimastix sp.…”

Section: Discussionmentioning

confidence: 99%

Cloning of a xylanase gene xyn2A from rumen fungus Neocallimastix sp. GMLF2 in Escherichia coli and its partial characterization

et al. 2009

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Anaerobic fungi belonging to the family Neocallimastigaceae are native inhabitants in the rumen of the most herbivores, such as cattle, sheep and goats. A member of this unique group, Neocallimastix sp. GMLF2 was isolated from cattle feces and screened for its xylanase encoding gene using polymerase chain reaction. The gene coding for a xylanase (xyn2A) was cloned in Escherichia coli and expression was monitored. To determine the enzyme activity, assays were conducted for both fungal xylanase and cloned xylanase (Xyl2A) for supernatant and cell-associated activities. Optimum pH and temperature of the enzyme were found to be 6.5 and 50• C, respectively. The enzyme was stable at 40• C and 50• C for 20 min but lost most of its activity when temperature reached 60• C for 5-min incubation time. Rumen fungal xylanase was mainly released to the supernatant of culture, while cloned xylanase activity was found as cell-associated. Multiple alignment of the amino acid sequences of Xyl2A with published xylanases from various organisms suggested that Xyl2A belongs to glycoside hydrolase family 11.

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“…The DNA sequences encoding AxeS20E and XynCDBFV were amplified by PCR from the plasmids pTriplEx2-S20E (Pai et al 2010a) and pNZJ021 (Liu et al 2005), respectively. The DNA sequences encoding the chimeric enzymes Axe-S2-Xyn and Xyn-S2-Axe were obtained by a modified overlap extension PCR method as described by Wurch et al (1998).…”

Section: Construction Of the Chimeric Enzymesmentioning

confidence: 99%

The Construction of Bifunctional Fusion Xylanolytic Enzymes and the Prediction of Optimum Reaction Conditions for the Enzyme Activity

Pai¹,

Wang²,

Guo³

et al. 2012

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Four chimeric xylanolytic enzymes were formed by fusion of a thermally stable xylanase XynCDBFV either to the N-terminus or C-terminus of a thermally stable acetylxylan esterase AxeS20E, with or without a Gly-rich flexible linker (S2). The three-dimensional (3D) structures of the chimeric enzymes were predicted using the I-TASSER server, and the results indicated that the structures of Axe-S2-Xyn and Xyn-S2-Axe were more similar to the native structures than were those of Axe-Xyn and Xyn-Axe. Axe-S2-Xyn and Xyn-S2-Axe were expressed in Escherichia coli and purified by means of affinity chromatography. Response surface modeling (RSM), combined with central composite design (CCD) and regression analysis, was then employed to optimize the xylanase activities of the chimeric enzymes. Under the optimal conditions, Xyn-S2-Axe had greater hydrolytic activities on natural xylans and rice straw than did the parental enzymes. These results suggested that the chimeric enzyme Xyn-S2-Axe could be effective at hydrolyzing xylan in biomass and that it has potential to be used in a range of biotechnological applications.

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Expression of Rumen Microbial Fibrolytic Enzyme Genes in Probiotic Lactobacillus reuteri

Cited by 43 publications

References 39 publications

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Cloning of a xylanase gene xyn2A from rumen fungus Neocallimastix sp. GMLF2 in Escherichia coli and its partial characterization

The Construction of Bifunctional Fusion Xylanolytic Enzymes and the Prediction of Optimum Reaction Conditions for the Enzyme Activity

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