Production and Characterization of Cellobiohydrolase from a Novel Strain of Penicillium purpurogenum KJS506

Lee, Kyoung-Mi; Joo, Ah-Reum; Jeya, Marimuthu; Lee, Kyoung-Min; Moon, Hee-Jung; Lee, Jung-Kul

doi:10.1007/s12010-010-9013-1

Cited by 10 publications

(3 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The higher V max and k cat together with lower K m and higher catalytic efficiency (k cat /K m ) indicate that pNP-G5 is a better substrate. The LqCel7B K m value for pNP-G5 is within the range reported for other GH7 cellobiohydrolases (22)(23)(24)(25)(26). The inhibitory effect of cellobiose, which is commonly observed in GH7 enzymes from fungi (15,16,(26)(27)(28), on the hydrolytic activity of LqCel7B was investigated using pNP-G5 (Fig.…”

Section: Resultssupporting

confidence: 77%

Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance

Kern

McGeehan

Streeter

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

Nature uses a diversity of glycoside hydrolase (GH) enzymes to convert polysaccharides to sugars. As lignocellulosic biomass deconstruction for biofuel production remains costly, natural GH diversity offers a starting point for developing industrial enzymes, and fungal GH family 7 (GH7) cellobiohydrolases, in particular, provide significant hydrolytic potential in industrial mixtures. Recently, GH7 enzymes have been found in other kingdoms of life besides fungi, including in animals and protists. Here, we describe the in vivo spatial expression distribution, properties, and structure of a unique endogenous GH7 cellulase from an animal, the marine wood borer Limnoria quadripunctata (LqCel7B). RT-quantitative PCR and Western blot studies show that LqCel7B is expressed in the hepatopancreas and secreted into the gut for wood degradation. We produced recombinant LqCel7B, with which we demonstrate that LqCel7B is a cellobiohydrolase and obtained four high-resolution crystal structures. Based on a crystallographic and computational comparison of LqCel7B to the well-characterized Hypocrea jecorina GH7 cellobiohydrolase, LqCel7B exhibits an extended substrate-binding motif at the tunnel entrance, which may aid in substrate acquisition and processivity. Interestingly, LqCel7B exhibits striking surface charges relative to fungal GH7 enzymes, which likely results from evolution in marine environments. We demonstrate that LqCel7B stability and activity remain unchanged, or increase at high salt concentration, and that the L. quadripunctata GH mixture generally contains cellulolytic enzymes with highly acidic surface charge compared with enzymes derived from terrestrial microbes. Overall, this study suggests that marine cellulases offer significant potential for utilization in high-solids industrial biomass conversion processes.gribble | carbohydrate degrading enzymes

show abstract

Section: Resultssupporting

confidence: 77%

Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance

Kern

McGeehan

Streeter

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

100

View full text Add to dashboard Cite

show abstract

“…For example, Le et al purified a novel cellobiohydrolase from Penicillium decumbens with a specific activity of 1.9 U mg −1 against p NPC [ 32 ]. Lee et al purified a cellobiohydrolase from Penicillium purpurogenum with a specific activity of 10.8 U mg −1 against p NPC U mg −1 [ 33 ]. Bok et al [ 11 ] purified two enzymes (CelA and CelB) with both endo- and exoglucanase activities.…”

Section: Resultsmentioning

confidence: 99%

A multifunctional thermophilic glycoside hydrolase from Caldicellulosiruptor owensensis with potential applications in production of biofuels and biochemicals

Peng

et al. 2016

Biotechnol Biofuels

View full text Add to dashboard Cite

BackgroundThermophilic enzymes have attracted much attention for their advantages of high reaction velocity, exceptional thermostability, and decreased risk of contamination. Exploring efficient thermophilic glycoside hydrolases will accelerate the industrialization of biofuels and biochemicals.ResultsA multifunctional glycoside hydrolase (GH) CoGH1A, belonging to GH1 family with high activities of β-d-glucosidase, exoglucanase, β-d-xylosidase, β-d-galactosidase, and transgalactosylation, was cloned and expressed from the extremely thermophilic bacterium Caldicellulosiruptor owensensis. The enzyme exerts excellent thermostability by retaining 100 % activity after 12-h incubation at 75 °C. The catalytic coefficients (kcat/Km) of the enzyme against pNP-β-D-galactopyranoside, pNP-β-D-glucopyranoside, pNP-β-D-cellobioside, pNP-β-D-xylopyranoside, and cellobiose were, respectively, 7450.0, 2467.5, 1085.4, 90.9, and 137.3 mM−1 s−1. When CoGH1A was supplemented at the dosage of 20 Ucellobiose g−1 biomass for hydrolysis of the pretreated corn stover, comparing with the control, the glucose and xylose yields were, respectively, increased 37.9 and 42.1 %, indicating that the enzyme contributed not only for glucose but also for xylose release. The efficiencies of lactose decomposition and synthesis of galactooligosaccharides (GalOS) by CoGH1A were investigated at low (40 g L−1) and high (500 g L−1) initial lactose concentrations. At low lactose concentration, the time for decomposition of 83 % lactose was 10 min, which is much shorter than the reported 2–10 h for reaching such a decomposition rate. At high lactose concentration, after 50-min catalysis, the GalOS concentration reached 221 g L−1 with a productivity of 265.2 g L−1 h−1. This productivity is at least 12-fold higher than those reported in literature.ConclusionsThe multifunctional glycoside hydrolase CoGH1A has high capabilities in saccharification of lignocellulosic biomass, decomposition of lactose, and synthesis of galactooligosaccharides. It is a promising enzyme to be used for bioconversion of carbohydrates in industrial scale. In addition, the results of this study indicate that the extremely thermophilic bacteria are potential resources for screening highly efficient glycoside hydrolases for the production of biofuels and biochemicals.

show abstract

“…Although improved strains of P. funiculosum have been isolated for overproduction of native enzymes, no genetically modified strains has been reported. Genes coding cellobiohydrolase GH7 have been cloned and characterized from a variety of fungal sources, including Humicola grisea [41], Trichoderma reesei [38,35], Penicillium janthinellum [21], P. chrysosporium [6,9] and Aspergillus sp. (4,40).…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning and Expression of a Family 7 Cellobiohydrolase Gene Cbh1 From Penicillium Funiculosum Ncl1

Vanitha¹,

Meera²,

Ramani³

et al. 2011

Int J of Micr Res

View full text Add to dashboard Cite

A cellobiohydrolase gene cbh1 was cloned from a cellulolytic fungus, Penicillum funiculosum NCL1. Nucleotide sequencing of cbh1 gene revealed that this gene was 1590 bp length encoding a putative protein consisting of 529 amino acids. The deduced amino acid sequence showed that the predicted molecular mass of the CBHI was 54.9 kDa, and showed significant homology to glycoside hydrolase family 7 cellobiohydrolases. The cbh1 gene was cloned using pET30b and expressed in E. coli BL21 (DE3). The expression analysis of the recombinant E. coli BL21 (pETC7) revealed the production of cbh1 transcript; however, functional cellobiohydrolase could not be detected. Therefore, the cbh1 gene was sub-cloned into GST tagged expression vector pGEX4t-3. The GST tagged cellobiohydrolase was purified to homogeneity using affinity chromatography. The recombinant enzyme exhibited optimum catalytic activity at pH 5.0 and 50 °C respectively. It was thermostable at 50 °C and retained 70% of its original activity after 30 min at 60 °C.

show abstract

Production and Characterization of Cellobiohydrolase from a Novel Strain of Penicillium purpurogenum KJS506

Cited by 10 publications

References 37 publications

Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance

Structural characterization of a unique marine animal family 7 cellobiohydrolase suggests a mechanism of cellulase salt tolerance

A multifunctional thermophilic glycoside hydrolase from Caldicellulosiruptor owensensis with potential applications in production of biofuels and biochemicals

Molecular Cloning and Expression of a Family 7 Cellobiohydrolase Gene Cbh1 From Penicillium Funiculosum Ncl1

Contact Info

Product

Resources

About