Skip to main content

Advertisement

SpringerLink
Log in

A cold-adapted endoglucanase from camel rumen with high catalytic activity at moderate and low temperatures: an anomaly of truly cold-adapted evolution in a mesophilic environment

  • Original Paper
  • Published:
Extremophiles Aims and scope Submit manuscript

Abstract

Endoglucanases are important enzymes in plant biomass degradation. They have current and potential applications in various industrial sectors including human and animal food processing, textile, paper, and renewable biofuel production. It is assumed that the cold-active endoglucanases, with high catalytic rates in moderate and cold temperatures, can improve the cost-effectiveness of industrial processes by lowering the need for heating and, thus, energy consumption. In this study, the endoglucanase CelCM3 was procured from a camel rumen metagenome via gene cloning and expression in Escherichia coli BL21 (DE3). The maximum activity of the enzyme on carboxymethyl cellulose (CMC) was obtained at pH 5 and 30 °C with a Vmax and Km of 339 U/mg and 2.57 mg/ml, respectively. The enzyme with an estimated low melting temperature of 45 °C and about 50% activity at 4 °C was identified to be cold-adapted. A thermodynamic analysis corroborated that CelCM3 with an activation energy (Ea), enthalpy of activation (ΔH), and Gibb’s free energy (ΔG) of, respectively, 18.47 kJ mol−1, 16.12 kJ mol−1, and 56.09 kJ mol−1 is a cold-active endoglucanase. In addition, CelCM3 was tolerant of metal ions, non-ionic detergents, urea, and organic solvents. Given these interesting characteristics, CelCM3 shows promise to meet the requirements of industrial applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Alvarez TM, Paiva JH, Ruiz DM, Cairo JPL, Pereira IO, Paixão DA, de Almeida RF, Tonoli CC, Ruller R, Santos CR (2013) Structure and function of a novel cellulase 5 from sugarcane soil metagenome. PLoS One 8(12):e83635

    Article  PubMed  PubMed Central  Google Scholar 

  • Bai X, Yua X, Wen A, Li J, Bai Y, Shao T (2016) Cloning, expression and characterization of a cold-adapted endo-1, 4-glucanase from Citrobacter farmeri A1, a symbiotic bacterium of Reticulitermes labralis. PeerJ 4:e2679

    Article  PubMed  PubMed Central  Google Scholar 

  • Bhat A, Riyaz-Ul-Hassan S, Ahmad N, Srivastava N, Johri S (2013) Isolation of cold-active, acidic endocellulase from Ladakh soil by functional metagenomics. Extremophiles 17:229–239

    Article  CAS  PubMed  Google Scholar 

  • Cavicchioli R, Charlton T, Ertan H, Omar SM, Siddiqui KS, Williams TJ (2011) Biotechnological uses of enzymes from psychrophiles. Microb Biotechnol 4(4):449–460

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cheng J, Huang S, Jiang H, Zhang Y, Li L, Wang J, Fan C (2016) Isolation and characterization of a non-specific endoglucanase from a metagenomic library of goat rumen. World J Microbiol Biotechnol 32:12

    Article  PubMed  Google Scholar 

  • Corrêa APF, Daroit DJ, Brandelli A (2010) Characterization of a keratinase produced by Bacillus sp. P7 isolated from an Amazonian environment. Int Biodeterior Biodegrad 64(1):1–6

    Article  Google Scholar 

  • Dadvar P, Mohammadabadi T, Sari M, Fayazi J (2016) Ruminal microbial fermentation of dromedary camel in a dual flow continuous culture system using cultivable and pasture forages. J Livest Sci Technol 4:17–24

    Google Scholar 

  • Dhar H, Kasana RC, Dutt S, Gulati A (2015a) Cloning and expression of low temperature active endoglucanase EG5C from Paenibacillus sp. IHB B 3084. Int J Biol Macromol 81:259–266

    Article  CAS  PubMed  Google Scholar 

  • Dhar H, Kasana RC, Gulati A (2015b) Heterologous expression and characterization of detergent stable endoglucanase EG5B from Paenibacillus sp. IHB B 3084. J Mol Catal B Enzym 120:9–15

    Article  CAS  Google Scholar 

  • Eriksson KE, Pettersson G (1968) Studies on cellulolytic enzymes: V. Some structural properties of the cellulase from Penicillium notatum. Arch Biochem Biophys 124:160–166

    Article  CAS  PubMed  Google Scholar 

  • Eyring H, Stearn AE (1939) The application of the theory of absolute reaction rates to proteins. Chem Rev 24(2):253–270

    Article  CAS  Google Scholar 

  • Feller G (2013) Psychrophilic enzymes: from folding to function and biotechnology. Scientifica 2013:1–28

    Article  Google Scholar 

  • Fu X, Liu P, Lin L, Hong Y, Huang X, Meng X, Liu Z (2010) A novel endoglucanase (Cel9P) from a marine bacterium Paenibacillus sp. BME-14. Appl Biochem Biotechnol 160(6):1627–1636

    Article  CAS  PubMed  Google Scholar 

  • Garsoux G, Lamotte J, Gerday C, Feller G (2004) Kinetic and structural optimization to catalysis at low temperatures in a psychrophilic cellulase from the Antarctic bacterium Pseudoalteromonas haloplanktis. Biochem J 384(2):247–253

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gharechahi J, Zahiri HS, Noghabi KA, Salekdeh GH (2015) In-depth diversity analysis of the bacterial community resident in the camel rumen. Syst Appl Microbiol 38(1):67–76

    Article  PubMed  Google Scholar 

  • Gunun P, Wanapat M, Anantasook N (2013) Rumen fermentation and performance of lactating dairy cows affected by physical forms and urea treatment of rice straw. Asian Australas J Anim Sci 26(9):1295–1303

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta V, Carrott P, Singh R, Chaudhary M, Kushwaha S (2016) Cellulose: a review as natural, modified and activated carbon adsorbent. Bioresour Technol 216:1066–1076

    Article  PubMed  Google Scholar 

  • Handelsman J (2004) Metagenomics: application of genomics to uncultured microorganisms. Microbiol Mol Biol Rev 68(4):669–685

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heinzelman P, Komor R, Kanaan A, Romero P, Yu X, Mohler S, Snow C, Arnold F (2010) Efficient screening of fungal cellobiohydrolase class I enzymes for thermostabilizing sequence blocks by SCHEMA structure-guided recombination. Protein Eng Des Sel 23(11):871–880

    Article  CAS  PubMed  Google Scholar 

  • Imran M, Anwar Z, Irshad M, Asad MJ, Ashfaq H (2016) Cellulase production from species of fungi and bacteria from agricultural wastes and its utilization in industry: a review. Adv Enzyme Res 4:44–55

    Article  CAS  Google Scholar 

  • Iqbal HMN, Ahmed I, Zia MA, Irfan M (2011) Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility. Adv Biosci Biotechnol 2:149–156

    Article  CAS  Google Scholar 

  • Jabbar A, Rashid MH, Javed MR, Perveen R, Malana MA (2008) Kinetics and thermodynamics of a novel endoglucanase (CMCase) from Gymnoascella citrina produced under solid-state condition. J Ind Microbiol Biotechnol 35(6):515–524

    Article  CAS  PubMed  Google Scholar 

  • Kasana RC, Gulati A (2011) Cellulases from psychrophilic microorganisms: a review. J Basic Microbiol 51:572–579

    Article  CAS  PubMed  Google Scholar 

  • Koppram R, Tomás-Pejó E, Xiros C, Olsson L (2014) Lignocellulosic ethanol production at high-gravity: challenges and perspectives. Trends Biotechnol 32(1):46–53

    Article  CAS  PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

    Article  CAS  PubMed  Google Scholar 

  • Liang C, Fioroni M, Rodríguez-Ropero F, Xue Y, Schwaneberg U, Ma Y (2011) Directed evolution of a thermophilic endoglucanase (Cel5A) into highly active Cel5A variants with an expanded temperature profile. J Biotechnol 154(1):46–53

    Article  CAS  PubMed  Google Scholar 

  • Martin M, Biver S, Steels S, Barbeyron T, Jam M, Portetelle D, Michel G, Vandenbol M (2014) Identification and characterization of a halotolerant, cold-active marine endo-1,4-glucanase by using functional metagenomics of seaweed-associated microbiota. Appl Environ Microbiol 80:4958–4967

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mathlouthi N, Mallet S, Saulnier L, Quemener B, Larbier M (2002) Effects of xylanase and β-glucanase addition on performance, nutrient digestibility, and physico-chemical conditions in the small intestine contents and caecal microflora of broiler chickens fed a wheat and barley-based diet. Anim Res 51(05):395–406

    Article  CAS  Google Scholar 

  • Mei HZ, Xia DG, Zhao QL, Zhang GZ, Qiu ZY, Qian P, Lu C (2016) Molecular cloning, expression, purification and characterization of a novel cellulase gene (Bh-EGaseI) in the beetle Batocera horsfieldi. Gene 576:45–51

    Article  CAS  PubMed  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Murashima K, Nishimura T, Nakamura Y, Koga J, Moriya T, Sumida N, Yaguchi T, Kono T (2002) Purification and characterization of new endo-1,4-β-d-glucanases from Rhizopus oryzae. Enzyme Microb Technol 30:319–326

    Article  CAS  Google Scholar 

  • Pang H, Zhang P, Duan C-J, Mo X-C, Tang J-L, Feng J-X (2009) Identification of cellulase genes from the metagenomes of compost soils and functional characterization of one novel endoglucanase. Curr Microbiol 58:404–408

    Article  CAS  PubMed  Google Scholar 

  • Pereira JH, Chen Z, McAndrew RP, Sapra R, Chhabra SR, Sale KL, Simmons BA, Adams PD (2010) Biochemical characterization and crystal structure of endoglucanase Cel5A from the hyperthermophilic Thermotoga maritima. J Struct Biol 172(3):372–379

    Article  CAS  PubMed  Google Scholar 

  • Rawat R, Kumar S, Chadha BS, Kumar D, Oberoi HS (2015) An acidothermophilic functionally active novel GH12 family endoglucanase from Aspergillus niger HO: purification, characterization and molecular interaction studies. Antonie Van Leeuwenhoek 107:103–117

    Article  CAS  PubMed  Google Scholar 

  • Samsudin AA, Wright ADG, Jassim RA (2012) Cellulolytic bacteria in the foregut of the dromedary camel (Camelus dromedarius). Appl Environ Microbiol 78(24):8836–8839

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sonan GK, Receveur-Brechot V, Duez C, Aghajari N, Czjzek M, Haser R, Gerday C (2007) The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. Biochem J 407(2):293–302

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tao YM, Xu XQ, Ma SJ, Liang G, Wu XB, Long MN, Chen QX (2011) Cellulase hydrolysis of rice straw and inactivation of endoglucanase in urea solution. J Agric Food Chem 59:10971–10975

    Article  CAS  PubMed  Google Scholar 

  • Tiwari R, Nain PKS, Singh S, Adak A, Saritha M, Rana S, Sharma A, Nain L (2015) Cold active holocellulase cocktail from Aspergillus niger SH3: process optimization for production and biomass hydrolysis. J Taiwan Inst Chem Eng 56:57–66

    Article  CAS  Google Scholar 

  • Ueda M, Ito A, Nakazawa M, Miyatake K, Sakaguchi M, Inouye K (2014) Cloning and expression of the cold-adapted endo-1,4-β-glucanase gene from Eisenia fetida. Carbohydr Polym 101:511–516

    Article  CAS  PubMed  Google Scholar 

  • Valaškova V, Baldrian P (2006) Degradation of cellulose and hemicelluloses by the brown rot fungus Piptoporus betulinus—production of extracellular enzymes and characterization of the major cellulases. Microbiology 152(12):3613–3622

    Article  PubMed  Google Scholar 

  • Vester JK, Glaring MA, Stougaard P (2014) Discovery of novel enzymes with industrial potential from a cold and alkaline environment by a combination of functional metagenomics and culturing. Microb Cell Fact 13(72):2–14

    Google Scholar 

  • Wilson DB (2011) Microbial diversity of cellulose hydrolysis. Curr Opin Microbiol 14:1–5

    Article  Google Scholar 

  • Yang J, Dang H (2011) Cloning and characterization of a novel cold-active endoglucanase establishing a new subfamily of glycosyl hydrolase family 5 from a psychrophilic deep-sea bacterium. FEMS Microbiol Lett 325(1):71–76

    Article  CAS  PubMed  Google Scholar 

  • Yennamalli RM, Rader AJ, Kenny AJ, Wolt JD, Sen TZ (2013) Endoglucanases: insights into thermostability for biofuel applications. Biotechnol Biofuels 6(1):136–144

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yin LJ, Lin HH, Xiao ZR (2010) Purification and characterization of a cellulase from Bacillus subtilis YJ1. J Mar Sci Technol 18:466–471

    Google Scholar 

  • Zarafeta D, Kissas D, Sayer C, Gudbergsdottir SR, Ladoukakis E, Isupov MN, Chatziioannou A, Peng X, Littlechild JA, Skretas G (2016) Discovery and characterization of a thermostable and highly halotolerant GH5 cellulase from an icelandic hot spring isolate. PLoS One 11(1):e0146454

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang G, Li S, Xue Y, Mao L, Ma Y (2012) Effects of salts on activity of halophilic cellulase with glucomannanase activity isolated from alkaliphilic and halophilic Bacillus sp. BG-CS10. Extremophiles 16(1):35–43

    Article  CAS  PubMed  Google Scholar 

  • Zhang F, Zhang XM, Yin YR, Li WJ (2015) Cloning, expression and characterization of a novel GH5 exo/endoglucanase of Thermobifida halotolerans YIM 90462 T by genome mining. J Biosci Bioeng 120(6):644–649

    Article  CAS  PubMed  Google Scholar 

  • Zhao S, Wang J, Zheng N, Bu D, Sun P, Yu Z (2015) Reducing microbial ureolytic activity in the rumen by immunization against urease therein. BMC Vet Res 11:94

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by National Institute of Genetic Engineering and Biotechnology (NIGEB) (Grant nos. 518 and 578).

Author information

Authors and Affiliations

  1. Department of Energy and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

    Kamran Khalili Ghadikolaei, Kambiz Akbari Noghabi & Hossein Shahbani Zahiri

  2. Human Genetics Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Javad Gharechahi

  3. Department of Plant Bioproducts, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Kamahldin Haghbeen

  4. Department of Systems Biology, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran

    Ghasem Hosseini Salekdeh

Authors
  1. Kamran Khalili Ghadikolaei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Javad Gharechahi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kamahldin Haghbeen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kambiz Akbari Noghabi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ghasem Hosseini Salekdeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hossein Shahbani Zahiri
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hossein Shahbani Zahiri.

Additional information

Communicated by L. Huang.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khalili Ghadikolaei, K., Gharechahi, J., Haghbeen, K. et al. A cold-adapted endoglucanase from camel rumen with high catalytic activity at moderate and low temperatures: an anomaly of truly cold-adapted evolution in a mesophilic environment. Extremophiles 22, 315–326 (2018). https://doi.org/10.1007/s00792-018-0999-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00792-018-0999-6

Keywords

Search

Navigation