A New Member of Family 11 Polysaccharide Lyase, Rhamnogalacturonan Lyase (CtRGLf) from Clostridium Thermocellum

Overview

Journal Mol Biotechnol

Publisher Springer

Specialties Biotechnology
Molecular Biology

Date 2016 Feb 28

PMID 26921189

Citations 4

Authors

Arun Dhillon

Vania O Fernandes

Fernando M V Dias

Jose A M Prates

Luis M A Ferreira

Carlos M G A Fontes

M S J Centeno

Arun Goyal

Affiliations

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Abstract

A thermostable, alkaline rhamnogalacturonan lyase (RG lyase) CtRGLf, of family 11 polysaccharide lyase from Clostridium thermocellum was cloned, expressed, purified and biochemically characterised. Both, the full-length CtRGLf (80 kDa) protein and its truncated derivative CtRGL (63.9 kDa) were expressed as soluble proteins and displayed maximum activity against rhamnogalacturonan I (RG I). CtRGLf showed maximum activity at 70 °C, while CtRGL at 60 °C. Both enzymes showed maximum activity at pH 8.5. CtRGLf and CtRGL do not show higher activity on substrates with high β-D-galactopyranose (D-Galp) substitution, this catalytic property deviates from that of some earlier characterised RG lyases which prefer substrates with high D-Galp substitution. The enzyme activity of CtRGLf and CtRGL was enhanced by 1.5 and 1.3 fold, respectively, in the presence of 3 mM of Ca(2+) ions. The TLC analysis of the degraded products of RG I, released by the action of CtRGLf and CtRGL revealed the production of RG oligosaccharides as major products confirming their endolytic activity.

Citing Articles

Composition and yield of non-cellulosic and cellulosic sugars in soluble and particulate fractions during consolidated bioprocessing of poplar biomass by Clostridium thermocellum.

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Biochemical Characterization of Two Rhamnogalacturonan Lyases From ATCC 8483 With Preference for RG-I Substrates.

Wang W, Wang Y, Yi H, Liu Y, Zhang G, Zhang L Front Microbiol. 2022; 12:799875.

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Molecular Cloning, Expression and Characterization of Pectin Methylesterase (CtPME) from Clostridium thermocellum.

Rajulapati V, Goyal A Mol Biotechnol. 2017; 59(4-5):128-140.

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Enzymatic diversity of the Clostridium thermocellum cellulosome is crucial for the degradation of crystalline cellulose and plant biomass.

Hirano K, Kurosaki M, Nihei S, Hasegawa H, Shinoda S, Haruki M Sci Rep. 2016; 6:35709.

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References

AMES G, Mimura C, Shyamala V . Bacterial periplasmic permeases belong to a family of transport proteins operating from Escherichia coli to human: Traffic ATPases. FEMS Microbiol Rev. 1990; 6(4):429-46. DOI: 10.1111/j.1574-6968.1990.tb04110.x. View

Ochiai A, Itoh T, Kawamata A, Hashimoto W, Murata K . Plant cell wall degradation by saprophytic Bacillus subtilis strains: gene clusters responsible for rhamnogalacturonan depolymerization. Appl Environ Microbiol. 2007; 73(12):3803-13. PMC: 1932723. DOI: 10.1128/AEM.00147-07. View

Pages S, Valette O, Abdou L, Belaich A, Belaich J . A rhamnogalacturonan lyase in the Clostridium cellulolyticum cellulosome. J Bacteriol. 2003; 185(16):4727-33. PMC: 166469. DOI: 10.1128/JB.185.16.4727-4733.2003. View

Anbar M, Gul O, Lamed R, Sezerman U, Bayer E . Improved thermostability of Clostridium thermocellum endoglucanase Cel8A by using consensus-guided mutagenesis. Appl Environ Microbiol. 2012; 78(9):3458-64. PMC: 3346478. DOI: 10.1128/AEM.07985-11. View

Koukiekolo R, Cho H, Kosugi A, Inui M, Yukawa H, Doi R . Degradation of corn fiber by Clostridium cellulovorans cellulases and hemicellulases and contribution of scaffolding protein CbpA. Appl Environ Microbiol. 2005; 71(7):3504-11. PMC: 1168997. DOI: 10.1128/AEM.71.7.3504-3511.2005. View

Silva I, Jers C, Otten H, Nyffenegger C, Larsen D, Derkx P . Design of thermostable rhamnogalacturonan lyase mutants from Bacillus licheniformis by combination of targeted single point mutations. Appl Microbiol Biotechnol. 2014; 98(10):4521-31. DOI: 10.1007/s00253-013-5483-8. View

Cheng H, Zhang Z, Leng J, Liu D, Hao M, Gao X . The inhibitory effects and mechanisms of rhamnogalacturonan I pectin from potato on HT-29 colon cancer cell proliferation and cell cycle progression. Int J Food Sci Nutr. 2012; 64(1):36-43. DOI: 10.3109/09637486.2012.694853. View

Das S, Ravindran R, Ahmed S, Das D, Goyal D, Fontes C . Bioethanol production involving recombinant C. thermocellum hydrolytic hemicellulase and fermentative microbes. Appl Biochem Biotechnol. 2012; 167(6):1475-88. DOI: 10.1007/s12010-012-9618-7. View

Oomen R, Doeswijk-Voragen C, Bush M, Vincken J, Borkhardt B, van den Broek L . In muro fragmentation of the rhamnogalacturonan I backbone in potato (Solanum tuberosum L.) results in a reduction and altered location of the galactan and arabinan side-chains and abnormal periderm development. Plant J. 2002; 30(4):403-13. DOI: 10.1046/j.1365-313x.2002.01296.x. View

10.

Moran F, Nasuno S, STARR M . Extracellular and intracellular polygllacturonic acid trans-eliminases of Erwinia carotovora. Arch Biochem Biophys. 1968; 123(2):298-306. DOI: 10.1016/0003-9861(68)90138-0. View

11.

Li S, Yang X, Bao M, Wu Y, Yu W, Han F . Family 13 carbohydrate-binding module of alginate lyase from Agarivorans sp. L11 enhances its catalytic efficiency and thermostability, and alters its substrate preference and product distribution. FEMS Microbiol Lett. 2015; 362(10). DOI: 10.1093/femsle/fnv054. View

12.

Lamed R, Setter E, Bayer E . Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum. J Bacteriol. 1983; 156(2):828-36. PMC: 217901. DOI: 10.1128/jb.156.2.828-836.1983. View

13.

Kashyap D, Vohra P, Chopra S, Tewari R . Applications of pectinases in the commercial sector: a review. Bioresour Technol. 2001; 77(3):215-27. DOI: 10.1016/s0960-8524(00)00118-8. View

14.

Laatu M, Condemine G . Rhamnogalacturonate lyase RhiE is secreted by the out system in Erwinia chrysanthemi. J Bacteriol. 2003; 185(5):1642-9. PMC: 148073. DOI: 10.1128/JB.185.5.1642-1649.2003. View

15.

McKie V, Vincken J, Voragen A, van den Broek L, Stimson E, Gilbert H . A new family of rhamnogalacturonan lyases contains an enzyme that binds to cellulose. Biochem J. 2001; 355(Pt 1):167-77. PMC: 1221724. DOI: 10.1042/0264-6021:3550167. View

16.

Lombard V, Ramulu H, Drula E, Coutinho P, Henrissat B . The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic Acids Res. 2013; 42(Database issue):D490-5. PMC: 3965031. DOI: 10.1093/nar/gkt1178. View

17.

Ridley B, ONeill M, Mohnen D . Pectins: structure, biosynthesis, and oligogalacturonide-related signaling. Phytochemistry. 2001; 57(6):929-67. DOI: 10.1016/s0031-9422(01)00113-3. View

18.

Vincken J, Schols H, Oomen R, McCann M, Ulvskov P, Voragen A . If homogalacturonan were a side chain of rhamnogalacturonan I. Implications for cell wall architecture. Plant Physiol. 2003; 132(4):1781-9. PMC: 1540329. DOI: 10.1104/pp.103.022350. View

19.

Olano-Martin E, Rimbach G, Gibson G, Rastall R . Pectin and pectic-oligosaccharides induce apoptosis in in vitro human colonic adenocarcinoma cells. Anticancer Res. 2003; 23(1A):341-6. View

20.

Matsunaga T, Ishii T, Matsumoto S, Higuchi M, Darvill A, Albersheim P . Occurrence of the primary cell wall polysaccharide rhamnogalacturonan II in pteridophytes, lycophytes, and bryophytes. Implications for the evolution of vascular plants. Plant Physiol. 2003; 134(1):339-51. PMC: 316313. DOI: 10.1104/pp.103.030072. View