Characterization of Extra Cellular Enzymes from Soil Actinomycetes: A Molecular Approach

International Journal of Biotech Trends and Technology (IJBTT)
© 2015 by IJBTT Journal
Volume - 5 Issue - 1                          
Year of Publication : 2015
Authors : Payal Das, Renu Solanki and Monisha Khanna

Payal Das, Renu Solanki and Monisha Khanna "Characterization of Extra Cellular Enzymes from Soil Actinomycetes: A Molecular Approach", International Journal of Biotech Trends and Technology (IJBTT), V5(1):20-32 Jan - Mar 2015, Published by Seventh Sense Research Group.


Extracellular enzymes from microbes are important biocatalysts with their widespread applications in several industries such as textile, biorefineries, food, pulp and paper, agriculture, detergent and pharmaceuticals. Actinomycetes are group of gram positive soil microbes, well known as producers of extracellular enzymes. Among the various genera of actinomycetes, Streptomyces has been a widely exploited group for production of these enzymes. Primary and secondary screening of isolates for identifying the highest enzyme producers followed by estimation of enzyme activity is the primary step. Immense work has been done for identification of genes, understanding the role of structural domains responsible for enzyme activity by X-ray crystallography, mass spectroscopy and NMR studies. This is followed by analyses of protein sequences, phylogenetic tree construction based on amino acid differences among species and recombinant studies for identification of protein families for hyper-producing the enzyme yield.


1. K. Haggag, A.A. Ragheb, I.A. EL-Thalouth, S.H. Nassar and H.E.L. Sayed, A review article on enzymes and their role in resist and discharge printing styles. Life Science Journal, 2013, Vol.10, p.1646-1654.
2. P. Baldrian, J. Šnajdr, V. Merhautova, P. Dobiašova, T. Cajthaml and Valaškova V, Responses of the extracellular enzyme activities in hardwood forest to soil temperature and seasonality and the potential effects of climate change. Soil Biol Biochem, 2013, Vol. 56, p. 60–68.
3. P. Baldrian, Distribution of extracellular enzymes in soils: spatial heterogeneity and determining factors at various scales. Soil Sci Soc Am J, 2014, Vol. 78, p.11–18.
4. R.G. Burns, J.L. DeForest, J. Marxsen, R.L. Sinsabaugh, M.E. Stromberger, M.D. Wallenstein, M.N. Weintraub and A. Zoppini, Soil enzymes in a changing environment: Current knowledge and future directions. Soil Biol Biochem, 2013, vol. 58, p. 216-234.
5. M. Sharma, Actinomycetes: source, identification, and their applications. Int J Curr Microbiol App Sci, 2014, vol. 3, p. 801-832.
6. K. Mojsov, Microbial cellulases and their applications in textile processing. International Journal of Marketing and Technology, 2012, vol. 2, p. 12-29.
7. S. Mohanram, D. Amat, J. Choudhary, A. Arora and L. Nai, Novel perspectives for evolving enzyme cocktails for lignocellulose hydrolysis in biorefineries. Sustainable Chemical Processes, 2013, Vol. 1, p. 15.
8. S.Y. Hwang, K. Nakashima, N. Okai, F. Okazaki, M. Miyake, K. Harazono, C. Ogino and A. Kondo, Thermal stability and starch degradation profile of α-amylase from Streptomyces avermitilis. Biosci Biotechnol Biochem, 2013, vol. 7, p. 2449-53.
9. N.A. Ibrahim, H.A. El-Shafei, M.S. Abdel-Aziz, M.F. Ghaly, B.M. Eid and A.A. Hamed, The potential use of alkaline protease from Streptomyces albidoflavus as an eco-friendly wool modifier. The Journal of the textile institute, 2012, vol. 103, p. 49-498.
10. H. Hamdali, M. Hafidi, M.J. Virolle and Y. Ouhdouch, Rock phosphate solubilizing actinomycetes: screening for plant growth-promoting activities. World J Microbiol Biotechnol, 2008, vol. 24, p. 2565-2575.
11. C.A. Oliveira, V.M.C Alves, I.E. Marriel, E.A. Gomes, M.R. Scotti, N.P. Carneiro, C.T. Guimara, R.E. Schaffert and N.M.H. Sa, Phosphate solubilizing microorganisms isolated from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome. Soil Biology and Biochemistry, 2009, vol. 41, p. 1782–1787.
12. G.M. Mohan and M.A.S. Charya, Enzymatic activity of freshwater actinomycetes. International Research Journal of Pharmacy, 2012, vol. 3, p. 193-197.
13. R. Solanki, M. Khanna and R. Lal, Review article entitled ―Bioactive compounds from marine actinomycetes. Indian Journal of Microbiology, 2008, vol. 48, p. 410-431.
14. M. Khanna, R. Solanki and R. Lal, Selective isolation of rare actinomycetes producing novel antimicrobial compounds. International Journal of advanced Biotechnology and Research, 2011, vol. 2, p.357- 375.
15. R. Solanki, R. Lal and M. Khanna, Antimicrobial activities of actinomycetes from diverse ecological habitats in Delhi and its adjoining states. Indian Journal of Microbial World, 2011, vol. 13, p.233-240.
16. P. Das, R. Solanki and M. Khanna, Isolation and screening of cellulolytic actinomycetes from diverse habitats. International Journal of Advanced Biotechnology and Research, 2014, vol. 5, p.438-451.
17. D.D. Teja, V.S. Devi, N. Harsha, S.S. Vishala and P.K.S. Lakshmi, Production of L-glutaminase from marine ecosystems and optimal conditions for maximal production by actinomycetes. International Journal of Advanced Research, 2014, vol. 2, p. 485-491.
18. P. Kumar, A. Kumar, T. Kumar and B.N. Singh, Purification of alkaline phosphatase (ALP) from Streptomyces sp Js-20 isolated from mangrove sediment. Asian J Exp Biol Sci, 2012, vol. 3, p.236 – 242.
19. S. Sadhu and T.K. Maiti, Cellulase production by bacteria: A review. British Microbiology Research Journal, 2013, vol. 3, p. 235-258.
20. P. Langan, A.K. Sangha, T. Wymore, J.M. Parks, Z.K. Yang, B.L. Hanson, Z. Fisher, S.A. Mason, P. Matthew, M.P. Blakeley, V.T. Forsyth, J.P. Glusker, H.L. Carrell, J.C. Smith, D.A. Keen, D.E. Graha and A. Kovalevsky, L-arabinose binding, isomerization, and epimerization by D-xylose isomerase: X-Ray/Neutron crystallographic and molecular simulation study. Structure, 2014, vol. 22, p.1287–1300.
21. C.S. Priya, N. Jagannathanv and P.T. Kalaichelvan, Production of chitinase by Streptomyces hygroscopicus VMCH2 by optimisation of cultural conditions. International Journal of Pharma and Bio Sciences, 2011, vol. 2, p.210-219
22. A.A. Al-Askar, Microbiological studies on the in vitro inhibitory effect of Streptomyces collinus albescens against some phytopathogenic fungi. African Journal of Microbiology Research, 2012, vol. 6, p.3277-3283.
23. I. Anderson, B. Abt, A. Lykidis, H.P. Klenk, N. Kyrpides and N. Ivanova, Genomics of aerobic cellulose utilization systems in actinobacteria. PLoS One, 2012, vol. 7, p.1-10.
24. M.R. Christopherson, G. Suen, S. Bramhacharya, K.A. Jewell, F.O. Aylward, D. Mead and P.J. Brumm, The genome sequences of Cellulomonas fimi and ―Cellvibrio gilvus reveal the cellulolytic strategies of two facultative anaerobes, transfer of ―Cellvibrio gilvus to the genus Cellulomonas, and proposal of Cellulomonas gilvus sp. nov. PLoS One, 2013, vol. 8, p.1-10.
25. R.E. Quiroz-Castañeda and J.L. Folch-Mallol, Sustainable Degradation of Lignocellulosic Biomass - Techniques, Applications and Commercialization, 2013,
26. C.Y. Chen, Y.C. Huang, C.M. Wei , M. Meng, W.H. Liu and C.H. Yang, Properties of the newly isolated extracellular thermo-alkali-stable laccase from thermophilic actinomycetes, Thermobifida fusca and its application in dye intermediates oxidation. AMB Express, 2013, vol. 3, p.1-9.
27. M.A.M. Yassien, A.A.M. Jiman-Fatani and H.Z. Asfour, Production, purification and characterization of cellulase from Streptomyces sp. African Journal of Microbiology, 2014, vol. 4, p.348-354.
28. A. Javmen, S. Grigiskis, M. Rudenkov and M. Mauricas, Purification and partial characterization of a novel β-1,3-endoglucanase from Streptomyces rutgersensis. Protein J, 2013, vol. 32, p.411-7.
29. M.N.F Cirigilano, R.C. Rezende, M.P.G Oliveira, P.H.F Pereira, R.P. Nascimento, E.P.S Bon, A. Macrae and R.R.R. Coelho, Streptomyces misionensis PESB-25 produces a thermoacidophilic endoglucanase using sugarcane bagasse and corn steep liquor as the sole organic substrates. BioMed Research International, 2013, p.1-9.
30. R.C. Kuhad, R. Gupta and A. Singh, Microbial cellulases and their industrial applications. Enzyme Research, 2011, p.1-10.
31. B. Azzeddine, M. Abdelaziz, C. Estelle, K. Mouloud, B. Nawel, B. Nabila, D. Francis and B. Said, Optimization and partial characterization of endoglucanase produced by Streptomyces sp. B-PNG23. Arch Biol Sci, 2013, vol. 65, p.549-558.
32. A. Shweta, Cellulases of bacterial origin and their applications: A review. International Journal of Science and Research, 2012, vol. 3, p.1652-1655.
33. Z.K. Bagewadi, A.G. Vernekar, A.Y. Patil, A.A. Limaye and V.M. Jain, Biodegradation of industrially important textile dyes by actinomycetes isolated from activated sludge. Biotechnol Bioinf Bioeng, 2011, vol. 1, p.351-360.
34. M. Sandgren, M. Wu, S. Karkehabadi, C. Mitchinson, B.R. Kelemen, E.A. Larenas, J. Stahlberg and H. Hansson, The structure of a bacterial cellobiohydrolase: The catalytic core of the Thermobifida fusca family GH6 cellobiohydrolase Cel6B. J Mol Biol, 2013, vol. 425, p.622–635.
35. G.J. Joo, Purification and characterization of an extracellular chitinase from the antifungal biocontrol agent Streptomyces halstedii. Biotechnol Letters, 2005, vol. 27, p. 1483-1486.
36. M.S. Brzezinska, U. Jankiewicz and A. Burkowska, Purification and characterization of Streptomyces albidoflavus antifungal components. Applied Biochemistry and Microbiology, 2013, vol. 49, p. 451–457.
37. S. Chakraborty, G. Raut, A. Khopade, K. Mahadik and C. Kokare, Study on calcium ion dependent α-amylase from haloalkaliphilic marine Streptomyces strain A3. Indian Journal of Biotechnology, 2012, vol. 11, p. 427-437.
38. A. Acharyabhatta, S.K. Kandula and R. Terli, Taxonomy and polyphasic characterization of alkaline amylase producing marine actinomycete Streptomyces rochei BTSS 1001. International Journal of Microbiology, 2013, p. 1-8.
39. S.S. Kanwar and S. Devi, Thermostable xylanases of microbial origin: Recent insights and biotechnological potential. The International Journal of Biotechnology, 2012, vol. 1, p.1-19.
40. P. Mander, Y.H. Choi, G.C. Pradeep, Y.S. Choi, J.H. Hong, S.S. Cho and J.C. Yoo, Biochemical characterization of xylanase produced from Streptomyces sp. CS624 using an agro residue substrate. Process Biochemistry, 2014, vol. 3, p. 451-456.
41. H.M. Rifaat, Z.A. Nahieb and Y.M. Ahmed, Production of xylanases by Streptomyces species and their bleaching effect on rice straw pulp. Applied Ecology and Environmental Research, 2005, vol. 4, p.151-160.
42. Z. Wang, Y. Jin, H. Wu, Z. Tian, Y. Wu and X. Xie, A novel, alkalitolerant thermostable xylanase from Saccharomonospora viridis: Direct gene cloning, expression and enzyme characterization. World J Microbiol Biotechnol, 2012, vol. 28, p.2741-8.
43. P. Manivasagan, J. Venkatesan, K. Sivakumar and S.K. Kim, Production, characterization and antioxidant potential of protease from Streptomyces sp. MAB18 using poultry wastes. BioMed Research International, 2013, p. 1-12.
44. K. Wateewuthajarn and P. Pinphanichakarn, Purification and characterization of xylanases from Streptomyces sp. PC22. J Sci Res Chula Univ, 2000, vol. 25, p.245-258.
45. B. Sowmya, D. Gomathi, M. Kalaiselvi, G. Ravikumar, C. Arulraj and C. Uma, Production and purification of chitinase by Streptomyces sp. from soil. J Adv Sci Res, 2012, vol. 3, p.25-29.
46. R. Kumar, A.L. Showkat, A. Sajad, P. Sasmita, K. Nitu, K. Devendra, N. Priyanka and Seenivasan, Cellulolytic activity of actinomycetes isolated from Areraj region, Bihar. International journal of current discoveries and innovations, 2013, vol. 2, p. 92-96.
47. Y.K. Mohanta, Isolation of cellulose degrading actinomycetes and evaluation of their cellulolytic potential. Bioengineering and Bioscience, 2014, vol. 2, p.1-5.
48. J. Ramakrishnan and M. Narayanan, Studies on xylanase producing thermophilic Streptomyces sp from compost soil. International Journal of Pharm Tech Research, 2013, vol. 5, p. 1386-1392.
49. R. Ghorbani-Nasrabadi, R. Greiner, H.A. Alikhani, J. Hamedi and B. Yakhchali, Distribution of actinomycetes in different soil ecosystems and effect of media composition on extracellular phosphatase activity. Journal of Soil Science and Plant Nutrition, 2013, vol. 13, p.223-236.
50. Y.A. Gherbawy, H. Elhariry, A. Altalhi and B. El-Deeb, Molecular screening of family 19 chitinase-producing Streptomyces and their potential antagonists against some phytopathogenic fungi. The Journal of Microbiology, 2012, vol. 50, p. 459–468.
51. P. Manivasagan, S. Gnanam, K. Sivakumar, T. Thangaradjou, S. Vijayalakshmi and T. Balasubramanian, Isolation, identification and characterization of multiple enzyme producing actinobacteria from sediment samples of Kodiyakarai coast, the Bay of Bengal. African Journal of Microbiology Research, 2010, vol. 4, p. 1550-1559.
52. T. Sriyapai, P. Somyoonsap, S. Areekit, P. Khawsak, A. Pakpitcharoen and K. Chansir, Isolation, cloning and molecular characterization of a thermotolerant xylanase from Streptomyces sp. THW31. African Journal of Biotechnology, 2013, vol. 12, p. 427-437.
53. R.S. Moura, J.F. Martin, A. Martin and P. Liras, Substrate analysis and molecular cloning of the extracellular alkaline phosphatase of Streptomyces griseus. Microbiology, 2001, vol. 147, p. 1525-1533.
54. A. Amore, O. Pepe, V. Ventorino, L. Birolo, C. Giangrande and V. Faraco, Cloning and recombinant expression of a cellulase from the cellulolytic strain Streptomyces sp. G12 isolated from compost. Microbial Cell Factories, 2012, vol. 11, p. 1-12.
55. N.E.A Naggar and N.A.M Abdelwahed, Optimization of process parameters for the production of alkali-tolerant carboxymethyl cellulase by newly isolated Streptomyces sp. strain NEAE-D. African Journal of Biotechnology, 2012, vol. 11, p.1185-1196.
56. F. A. Mansour, A.A Shereif, E.D.M.I.M. Nour, M.I. Abou-Dobara and A.S. Ball, Purification and characterization of xylanase from a thermophilic Streptomyces sp. K37. Acta Microbiologica Polonica, 2003, vol. 52, p.159-72.
57. G.C.P, H.Y. Yoo, S.S. Cho, Y.H. Choi and J.C. Yoo, An extracellular chitinase from Streptomyces sp. CS147 releases N-acetyl-Dglucosamine (GlcNAc) as principal product. Appl Biochem Biotechnol, 2015, vol. 175, p.372-86.
58. R. Ragunathan and R.P. Dhas, Exploitation of novel actinomycetes isolated from Western Ghats of Tamil Nadu and production of xylanase by Streptomyces spp. pds1. Science Journal of Microbiology, 2013, doi: 10.7237/sjmb/174.
59. A. Nagpure and R.K Gupta, Purification and characterization of an extracellular chitinase from antagonistic Streptomyces violaceusniger. J Basic Microbiol, 2013, vol. 53, p. 429-39.
60. S. Yano, N. Rattanakit, A. Honda, Y. Noda, M. Wakayama, A. Plikomol and T. Tachiki, Purification and characterization of chitinase A of Streptomyces cyaneus SP-27: an enzyme participates in protoplast formation from Schizophyllum commune mycelia. Bioscience Biotechnology and Biochemistry, 2008, vol.72, p.54-61.
61. I.R. Byman, Kinetic and functional charaterization of a family 19 chitinase from Streptomyces coelicolor A3 (2). Master’s Thesis Norwegian University of Life Sciences Ås, 2014,
62. K.C. Hoang, T.H. Lai, C.S. Lin, Y.T. Chen and C.Y. Liau, The Chitinolytic Activities of Streptomyces sp. TH-11. Int J Mol Sci, 2011, vol. 12, p. 56–65.
63. C.C. Chen, H. Luo, X. Han, P. Lv, T.P. Ko, W. Peng, C.H. Huang, K. Wang, J. Gao, Y. Zheng, Y.Yang, J. Zhang, B. Yao and R.T. Guo, Structural perspectives of an engineered β-1, 4-xylanase with enhanced thermostability. Journal of Biotechnology, 2014, vol. 189, p. 175–182.
64. I.A. Hoell, B. DaIhus, E.B. Heggset, S.I. Aspmo and V.G.H Eijsink, Crystal structure and enzymatic properties of a bacterial family 19 chitinase reveal differences from plant enzymes. FEBS Journal, 2006, vol. 273, p. 4889–4900.
65. H. Nacke, M. Engelhaupt, S. Brady, C. Fischer, J. Tautzt, Daniel R (2012) Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes. Biotechnol Lett 34:663-75.
66. M.P. Munar, Y.G. Eom, Y.S. Kim and T.J. Kim, Polymorphism in cellulase genes celB, celS2 and celS, distinguishes Streptomyces coelicolor M145 from unknown soil actinomycete isolates. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 2013, vol. 5, p.1908-1915.
67. Z. Liu, X. Zhao and F. Bai, Production of xylanase by an alkalinetolerant marine-derived Streptomyces viridochromogenes strain and improvement by ribosome engineering. Appl Microbiol Biotechnol, 2013, vol. 97, p.4361-8.
68. M. Diaz, S. Rodriguez, J.M. Fernández-Abalos, J.D.L Rivas, A. Ruiz- Arribas, V.L. Shnyrov and R.I. Santamaría, Single mutations of residues outside the active center of the xylanase Xys1Δ from Streptomyces halstedii JM8 affect its activity. FEMS Microbiology Letters, 2004, vol. 240, p. 237–243.
69. S. Datta, L.R. Christena and Y.R.S Rajaram, Enzyme immobilization: an overview on techniques and support materials. 3 Biotech, 2013, vol. 3, p.1–9.

Extracellular enzymes from actinomycetes, Primary and Secondary screening, Structural analysis by Xray crystallography and Mass spectroscopy, Gene and protein phylogeny, Recombinant studies.