Research Article | Volume: 6, Issue: 3, May-June, 2018
Morphological, enzymatic screening, and phylogenetic analysis of thermophilic bacilli isolated from five hot springs of Myagdi, Nepal
The present study was conducted to identify and characterize the thermophilic bacteria isolated from five hot springs, namely, Sinkosh, Singha, Bhurung, Ratopani, and Paudwar located in Myagdi district, Nepal, using phenotypic and genotypic methods. The hot spring has temperature 42–62°C and pH 6.5–6.8. Isolation of thermophiles was done using simple enriched nutrient broth media at 60°C. Selected strains were screened for thermostable enzymes; cellulase, hemicellulase, amylase, protease, gelatinase, and lipase using substrates carboxymethylcellulose, xylan, soluble starch, casein, gelatin, and Tween (20, 40, 60, or 80), respectively. The bacteria were grouped into 16 groups based on morphological and biochemical characteristics. 16S rRNA sequence analysis of 16 isolates and phylogenetic analysis showed a cluster of five distinct taxonomic groups. The groups were identified as genus Anoxybacillus, Aeribacillus, Brevibacillus, Bacillus, and Geobacillus, based on ≥95% similarity with reference strains. This is the first study that reports Anoxybacillus sp., Brevibacillus sp., and Aeribacillus sp. from the hot springs of Nepal.
Yadav P, Korpole S, Prasad GS, Sahni G, Maharjan J, Sreerama L, Bhattarai T. Morphological, enzymatic screening, and phylogenetic analysis of thermophilic bacilli isolated from five hot springs of Myagdi, Nepal. J App Biol Biotech. 2018;6(03):1-8.
1. Averhoff B, Müller V. Exploring research frontiers in microbiology: recent advances in halophilic and thermophilic extremophiles. Research in microbiology. 2010;161(6):506-14. https://doi.org/10.1016/j.resmic.2010.05.006 | |
2. Meyerâ€Dombard D, Shock E, Amend J. Archaeal and bacterial communities in geochemically diverse hot springs of Yellowstone National Park, USA. Geobiology. 2005;3(3):211-27. https://doi.org/10.1111/j.1472-4669.2005.00052.x | |
3. Aanniz T, Ouadghiri M, Melloul M, Swings J, Elfahime E, Ibijbijen J, et al. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils. Brazilian Journal of Microbiology. 2015;46(2):443-53. https://doi.org/10.1590/S1517-838246220140219 | |
4. Badhai J, Ghosh TS, Das SK. Taxonomic and functional characteristics of microbial communities and their correlation with physicochemical properties of four geothermal springs in Odisha, India. Frontiers in microbiology. 2015;6. https://doi.org/10.3389/fmicb.2015.01166 | |
5. Bisht SS, Panda AK. Biochemical characterization and 16S rRNA sequencing of few lipase-producing thermophilic bacteria from Taptapani hot water spring, Orissa, India. Biotechnology research international. 2011;2011. | |
6. Gray KA, Zhao L, Emptage M. Bioethanol. Current opinion in chemical biology. 2006;10(2):141-6. https://doi.org/10.1016/j.cbpa.2006.02.035 | |
7. Wyman CE. Potential synergies and challenges in refining cellulosic biomass to fuels, chemicals, and power. Biotechnology progress. 2003;19(2):254-62. https://doi.org/10.1021/bp025654l | |
8. Zaldivar J, Nielsen J, Olsson L. Fuel ethanol production from lignocellulose: a challenge for metabolic engineering and process integration. Applied microbiology and biotechnology. 2001;56(1):17-34. https://doi.org/10.1007/s002530100624 | |
9. Rabinovich M. ethanol production from materials containing cellulose: the potential of approaches developed in Russia. Prikladnaia biokhimiia i mikrobiologiia. 2006;42(1):5-32. | |
10. Ranjit M, editor Geothermal energy update of Nepal. Proceedings of the 2010 World Geothermal Congress; 2010. | |
11. Yadav RKP, Timilsina A, Yadawa RK, Pokhrel CP. Potential Cellulosic Ethanol Production from Organic Residues of Agro-Based Industries in Nepal. ISRN Renewable Energy. 2014;2014:6. https://doi.org/10.1155/2014/305695 | |
12. Akmar HN, Asma I, Venugopal B, Latha LY, Sasidharan S. Identification of appropriate sample and culture method for isolation of new thermophilic bacteria from hot spring. African Journal of Microbiology Research. 2011;5(3):217-21. | |
13. Silva Pd, Nahas E. Bacterial diversity in soil in response to different plans, phosphate fertilizers and liming. Brazilian Journal of Microbiology. 2002;33:304-10. https://doi.org/10.1590/S1517-83822002000400005 | |
14. Mc Faddin. J.F. Pruebas bioquimicas para la identificacion de bacterias de importancia clinica. . Panamericana, Buenos Aires. 1984: 11-301. | |
15. Abd-Elhalem BT, El-Sawy M, Gamal RF, Abou-Taleb KA. Production of amylases from Bacillus amyloliquefaciens under submerged fermentation using some agro-industrial by-products. Annals of Agricultural Sciences. 2015;60(2):193-202. https://doi.org/10.1016/j.aoas.2015.06.001 | |
16. Zhou M-Y, Chen X-L, Zhao H-L, Dang H-Y, Luan X-W, Zhang X-Y, et al. Diversity of both the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea. Microbial ecology. 2009;58(3):582-90. https://doi.org/10.1007/s00248-009-9506-z | |
17. Kasana RC, Salwan R, Dhar H, Dutt S, Gulati A. A rapid and easy method for the detection of microbial cellulases on agar plates using Gram's iodine. Current microbiology. 2008;57(5):503-7. https://doi.org/10.1007/s00284-008-9276-8 | |
18. Singh AK, Tripathi B, Sahay H, Singh R, Kaushik R, Saxena A, et al. Biochemical and molecular characterization of thermo-alkali tolerant xylanase producing bacteria from thermal springs of Manikaran. Indian journal of microbiology. 2010;50:2-9. https://doi.org/10.1007/s12088-010-0071-4 | |
19. Tanasupawat S, Phoottosavako M, Keeratipibul S. Characterization and lipolytic activity of lactic acid bacteria isolated from Thai fermented meat. 2015. | |
20. Whaley D, Dowell V, Wanderlinder L, Lombard G. Gelatin agar medium for detecting gelatinase production by anaerobic bacteria. Journal of clinical microbiology. 1982;16(2):224-9. | |
21. Padilha I, Carvalho L, Dias P, Grisi T, Silva F, Santos S, et al. Production and characterization of thermophilic carboxymethyl cellulase synthesized by Bacillus sp. Growing on sugarcane bagasse in submerged fermentation. Brazilian Journal of Chemical Engineering. 2015;32(1):35-42. https://doi.org/10.1590/0104-6632.20150321s00003298 | |
22. Assareh R, Zahiri HS, Noghabi KA, Aminzadeh S. Characterization of the newly isolated Geobacillus sp. T1, the efficient cellulase-producer on untreated barley and wheat straws. Bioresource technology. 2012;120:99-105. https://doi.org/10.1016/j.biortech.2012.06.027 | |
23. Yassien MA-M, Jiman-Fatani AAM, Asfour HZ. Production, purification and characterization of cellulase from Streptomyces sp. African Journal of Microbiology Research. 2014;8(4):348-54. https://doi.org/10.5897/AJMR2013.6500 | |
24. Bhalla A, Bischoff KM, Sani RK. Highly thermostable xylanase production from a thermophilic Geobacillus sp. strain WsUcF1 utilizing lignocellulosic biomass. Frontiers in bioengineering and biotechnology. 2015;3. https://doi.org/10.3389/fbioe.2015.00084 | |
25. Rasooli I, Astaneh SDA, Borna H, Barchini KA. A thermostable α-amylase producing natural variant of Bacillus spp. isolated from soil in Iran. Am J Agric Biol Sci. 2008;3(3):591-6. https://doi.org/10.3844/ajabssp.2008.591.596 | |
26. Lo Y-C, Saratale GD, Chen W-M, Bai M-D, Chang J-S. Isolation of cellulose-hydrolytic bacteria and applications of the cellulolytic enzymes for cellulosic biohydrogen production. Enzyme and Microbial Technology. 2009;44(6):417-25. https://doi.org/10.1016/j.enzmictec.2009.03.002 | |
27. Bond PL, Smriga SP, Banfield JF. Phylogeny of microorganisms populating a thick, subaerial, predominantly lithotrophic biofilm at an extreme acid mine drainage site. Applied and Environmental Microbiology. 2000;66(9):3842-9. https://doi.org/10.1128/AEM.66.9.3842-3849.2000 | |
28. Singh PK, Kumari A, Chawla N, Pinnaka AK, Korpole S. Rhodococcus lactis sp. nov., an actinobacterium isolated from sludge of a dairy waste treatment plant. International journal of systematic and evolutionary microbiology. 2015;65(11):4215-20. https://doi.org/10.1099/ijsem.0.000565 | |
29. Kim O-S, Cho Y-J, Lee K, Yoon S-H, Kim M, Na H, et al. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. International journal of systematic and evolutionary microbiology. 2012;62(3):716-21. https://doi.org/10.1099/ijs.0.038075-0 | |
30. Kamble RD, Jadhav AR. Isolation, purification, and characterization of xylanase produced by a new species of Bacillus in solid state fermentation. International Journal of Microbiology. 2012;2012. | |
31. Harish Adhikari, Sangam Ghimire, YK BK. Enzymatic screening and molecular characterization of thermophilic bacterial strains isolated from hotspring of tatopani, bhurung, nepal. International Journal of Applied Sciences. 2015;3:392-7. https://doi.org/10.3126/ijasbt.v3i3.12724 | |
32. Kristjansson JK. Thermophilic organisms as sources of thermostable enzymes. Trends in Biotechnology. 1989;7(12):349-53. https://doi.org/10.1016/0167-7799(89)90035-8 | |
33. Liang Y, Feng Z, Yesuf J, Blackburn JW. Optimization of growth medium and enzyme assay conditions for crude cellulases produced by a novel thermophilic and cellulolytic bacterium, Anoxybacillus sp. 527. Applied biochemistry and biotechnology. 2010;160(6):1841-52. https://doi.org/10.1007/s12010-009-8677-x | |
34. Goh KM, Kahar UM, Chai YY, Chong CS, Chai KP, Ranjani V, et al. Recent discoveries and applications of Anoxybacillus. Applied microbiology and biotechnology. 2013;97(4):1475-88. https://doi.org/10.1007/s00253-012-4663-2 | |
35. Lynd LR, Weimer PJ, Van Zyl WH, Pretorius IS. Microbial cellulose utilization: fundamentals and biotechnology. Microbiology and molecular biology reviews. 2002;66(3):506-77. https://doi.org/10.1128/MMBR.66.3.506-577.2002 | |
36. Logan N, Berkeley R. Identification of Bacillus strains using the API system. Microbiology. 1984;130(7):1871-82. https://doi.org/10.1099/00221287-130-7-1871 | |
37. Yohandini H. Isolation and Phylogenetic Analysis of Thermophile Community Within Tanjung Sakti Hot Spring, South Sumatera, Indonesia. HAYATI Journal of Biosciences. 2015;22(3):143-8. https://doi.org/10.1016/j.hjb.2015.10.006 | |
38. Aditiawati P, Yohandini H, Fida Madayanti A. Microbial diversity of acidic hot spring (Kawah Hujan B) in geothermal field of Kamojang area, west Java-Indonesia. The open microbiology journal. 2009;3:58. https://doi.org/10.2174/1874285800903010058 | |
39. Cihan AC, Tekin N, Ozcan B, Cokmus C. The genetic diversity of genus Bacillus and the related genera revealed by 16S rRNA gene sequences and ardra analyses isolated from geothermal regions of turkey. Brazilian Journal of Microbiology. 2012;43(1):309-24. https://doi.org/10.1590/S1517-83822012000100037 | |
40. Baker GC, Gaffar S, Cowan DA, Suharto AR. Bacterial community analysis of Indonesian hot springs. FEMS Microbiology Letters. 2001;200(1):103-9. https://doi.org/10.1111/j.1574-6968.2001.tb10700.x | |
41. Turner P, Mamo G, Karlsson EN. Potential and utilization of thermophiles and thermostable enzymes in biorefining. Microbial cell factories. 2007;6(1):9. https://doi.org/10.1186/1475-2859-6-9 | |
42. Paes G, O'Donohue MJ. Engineering increased thermostability in the thermostable GH-11 xylanase from Thermobacillus xylanilyticus. Journal of biotechnology. 2006;125(3):338-50. https://doi.org/10.1016/j.jbiotec.2006.03.025 | |
43. Yan S, Wu G. Secretory pathway of cellulase: a mini-review. Biotechnology for biofuels. 2013;6(1):177. https://doi.org/10.1186/1754-6834-6-177 | |
44. Kahar UM, Chan K-G, Salleh MM, Hii SM, Goh KM. A high molecular-mass Anoxybacillus sp. SK3-4 amylopullulanase: characterization and its relationship in carbohydrate utilization. International journal of molecular sciences. 2013;14(6):11302-18. https://doi.org/10.3390/ijms140611302 | |
45. Tsuda T, Sasaki Y, Kawashima R. Physiological Aspects of Digestion and Metabolism in Ruminants: Proceedings of the Seventh International Symposium on Ruminant Physiology: Academic Press; 2012. | |
46. Rastogi G, Muppidi GL, Gurram RN, Adhikari A, Bischoff KM, Hughes SR, et al. Isolation and characterization of cellulose-degrading bacteria from the deep subsurface of the Homestake gold mine, Lead, South Dakota, USA. Journal of industrial microbiology & biotechnology. 2009;36(4):585. https://doi.org/10.1007/s10295-009-0528-9 | |
47. Logan NA, Forsyth G, Lebbe L, Goris J, Heyndrickx M, Balcaen A, et al. Polyphasic identification of Bacillus and Brevibacillus strains from clinical, dairy and industrial specimens and proposal of Brevibacillus invocatus sp. nov. International journal of systematic and evolutionary microbiology. 2002;52(3):953-66. | |
Year
Month
The Characterization of Amylolytic Enzyme Present in Fermented Sweet Sap of Palmyrah
Arumugam Vengadaramana, Mehala Uthayasooriyan, Thananthika Sittampalam, Nirosha Razeek, Ranganathan KapilanExpression analysis of recombinant Vigna radiata plant defensin 1 protein in transgenic tobacco plants
Hoang Thi Thao , Nguyen Thi Ngoc Lan, Ho Manh Tuong, Nguyen Vu Thanh Thanh, Le Van Son, Chu Hoang MauIn Vitro Antidiabetic and Anti-oxidant Activities of Methanol Extract of Tinospora Sinensis
Anindita Banerjee, Bithin Maji, Sandip Mukherjee, Kausik Chaudhuri, Tapan SealCharacterization of extracellular amylase from Bacillus sp. strain RU1
Aleem Basha Pinjari, Vijayalakshmi KotariScreening and identification of amylase producing strains of Bacillus
Kumar Pranay, Shree Ram Padmadeo, Vijay Jha, Birendra PrasadIn vitro anti-amylase activity of some Indian dietary spices
Bhosale Hemlata, Gawali Pornima, Kadam Tukaram, Baisthakur PankajSequestration and purification of essential enzyme from Pleurotus florida compost waste and its application studies
S Karthika Devi, N Prabhu , S Monika, N M Swapna, S Vaishali, T GajendranBioprospecting for extracellular enzymes from endophytic bacteria isolated from Vigna radiata and Cajanus cajan
Namita Bhutani, Rajat Maheshwari, Pradeep Kumar, Rakhi Dahiya, Pooja SunejaMolecular docking of active compounds from Phoenix pusilla root extract against antidiabetic and anti-inflammatory drug targets
Vijayabharathi Saravanan, Anuradha Venkatraman, Habeeb Shaik MohideenOptimization of amylase and protease production from oyster mushrooms koji (Pleurotus spp.) using response surface methodology
Nguyen Thi Ngoc Giang, Tran Van Khai, Nguyen Minh ThuyIsolation and partial characterization of amylase produced by fungal isolates from the agro-industrial waste source
Soumya Nandi, Twinkle Bose, Susmita Mahato, Somashree Chatterjee, Annalakshmi ChatterjeeIsolation and characterization of starch degrading bacteria from disparate soil samples
Vidhyutha Srivathsan, Mahima Bhandari, Priya SwaminathanCharacterization of an alkaline endo-α-1-4-amylase from alkaliphilic Bacillus halodurans LBW 5117 and demonstration of its desizing potential
Kevin Raymond Oluoch, Edward Kirwa Muge, Yvonne Wanjiku Mwangi, Francis Jakim MulaaA low-cost production, characterization, and application of raw starch degrading enzyme from the thermophilic filamentous bacterium, Laceyella sacchari P43
Wanthanee Khetkorn, Ausawadee Phonlamai, Chatsuda Sakdapetsiri, Vichien Kitpreechavanich, Thanasak LomthongBioconversion of brewer’s rice into ethanol by Saccharomyces cerevisiae using commercial amylases
Chandrasekhar Banoth,, Bhima BhukyaBioconversion of waste newspaper into fermentable sugars at different temperatures with different Aspergillus niger cellulase concentrations
J Boitumelo M Sibiya, J Pieter H van WykBiocatalysis of agro-processing waste by marine Streptomyces fungicidicus strain RPBS-A4 for cellulase production
Rajanikanth Akurathi, Damodharam ThotiProduction and purification of extracellular fungal cellulases using agricultural waste
Abishna Burugu, Dheerendra Kumar Suman, Chandrasekhar ChandaOptimization and statistical modeling of microbial cellulase production using submerged culture
Pratibha Maravi, Anil KumarIsolation and identification of bacteria with cellulose-degrading potential from soil and optimization of cellulase production
Shweta Ashok Bhagat, Seema Sambhaji KokitkarUpsurge production of cellulase from maize stover under soildstate conditions mediated by Streptomyces enissocaesilis DQ026641
Ashok Sudarshan, Siddanna Renuka, Sirasagar Reshma, Bhalerao Shilanjali, Dayanand AgsarSolid-state fermentation of groundnut (Arachis hypogaea) shell using Trichoderma sp., tape yeast, and tempeh yeast to produce cellulase
Muhammad Yusuf Abduh, Chalil Rizqullah Ramadhan, Alfanny Putri Fadhlilah, Siti Dhiffah Nabilah Abdul, Khairul Hadi BurhanIsolation and Characterization of Cellulase-Producing Myxobacterial Strain from the Unique Niche of Mirgund Wetland from the North-Western Himalayas
Daljeet Singh Dhanjal, Simranjeet Singh, Vijay Kumar, Praveen C. Ramamurthy, Chirag Chopra, Atif Khurshid Wani, Reena Singh, Joginder SinghIsolation and screening of potential lignocellulolytic microbes from Phra Nakhon Si Ayutthaya Province
Sunisa Suwannaphan, Paweena Suksaard, Piyangkun Lueangjaroenkit, Suvichark Aroonluk, Phakhwan ThongrakMutualistic synergy between Eudrilus eugeniae gut microbiota and Beta vulgaris root endophytes in organic matter decay
Pushpa Reddy, T. K. Suma, Amlan Hossain, Sudarsan SarkarMutational enhancement of Aspergillus niger Tiegh. for higher cellulase production comparable to Trichoderma species in solid-state fermentation
Harjeet Singh, Komal Janiyani, Ajit GangawaneAlbizia lebbeck fruit pods as a substrate for the production of lignocellulolytic enzymes by Aspergillus niger
Mamatha Pingili, Shailaja Raj Marla, Ramakrishna RaparlaProduction and optimization of enzyme xylanase by Aspergillus flavus using agricultural waste residues
Jyoti Richhariya, Tirthesh Kumar Sharma, Sippy DassaniOptimization of process and conditions for enhanced xylanase production under SSF using inexpensive agro-industrial waste
Vimalashanmugam Kanagasabai, Karuppaiya MaruthaiCloning and expression of a GH11 xylanase from Bacillus pumilus SSP-34 in Pichia pastoris GS115: Purification and characterization
Sagar Krishna Bhat,, Kavya Purushothaman, Appu Rao Gopala Rao Appu Rao, K Ramachandra KiniEngineering to enhance thermostability of xylanase: For the new era of biotechnology
Chitranshu Pandey, Pallavi Sharma, Neeraj GuptaFermentation medium optimization for the 1,4-ß-Endoxylanase production from Bacillus pumilus using agro-industrial waste
Varsha D. Savanth, B. S. Gowrishankar, K. B. RoopaIsolation and identification of pathogenic microbes from tomato puree and their delineation of distinctness by molecular techniques
R.K. Garg, N. Batav, N. Silawat, R.K. SinghIsolation and Identification by Morphological and Biochemical Methods of Antibiotic Producing Microorganisms from the gut of Macrotermes michaelseni in Maseno, Kenya
Aswani Susan Ayitso, David Miruka OnyangoModulation of some biochemical complications arising from alloxan-induced diabetic conditions in rats treated with Senna occidentalis leaf extract
Ojochenemi Ejeh Yakubu, Okwesili Fred Chiletugo Nwodo, Chinedu Imo, Sylvester Michael Chukwukadibia Udeh, Mikailu Abdulrahaman, Maryval Ogaku OgriBiochemical Modulations in Duttaphrynus melanostictus Tadpoles, Following Exposure to Commercial Formulations of Cypermethrin: An Overlooked Impact of Extensive Cypermethrin use
David Muniswamy, Shrinivas S Jadhav, Kartheek R MalowadeSubmerged fermentation, purification, and characterization of L-asparaginase from Streptomyces sp. isolated from soil
Savitha S. Desai, Basavaraj S. HungundConsequence of chromium-tainted soil on physical and biochemical responses of Vigna radiata L.
Bibhu Prasad Rath, Sujata Hota, Subhra Subhadarshini, Debasis Dash, Prabhu Kaibalya DasEffect of frozen storage on the biochemical, microbial and sensory attributes of Skipjack Tuna (Katsuwonus pelamis) fish loins
Suprakash Chakma, Susmita Saha, Nazmul Hossain, Md. Arifur Rahman, Marjan Akter, Md. Sazedul Hoque, Md. Rahamat Ullah, Sujan Kanti Mali, Al ShahriarBioprospecting of endophytic bacteria from the Indian Himalayas and their role in plant growth promotion of maize (Zea mays L.)
Kusam Lata Rana, Divjot Kour, Tanvir Kaur, Rubee Devi, Ashok Yadav, Ajar Nath YadavDengue infection profile in Gujarat, West India: A recent report
Nishita S. Naik, Shiva V. Murarka, Dhariya Sanghrajka, Bhavini S. Shah, Rutvik J. Raval, Mandava V. RaoSeasonal analysis of certain biochemical parameters of carps cultured in domestic sewage oxidation ponds
Sharique A. Ali, M. Hanumanth Raju, Naima ParveenAllelopathic potential of Annona muricata (L.) on physiological and biochemical changes of Vigna radiata (L.) and Eleusine coracana (L.) Gaertn.
Edwina Kannan, Leela PalayianCalcium influences biochemical and antioxidant enzymatic activities in tomato fruits during storage
Salma Santhosh, Thiyagarajan Chitdeshwari, Kavitha ChinnasamyRole of effective microorganisms on hematological and biochemical indices of cultured Oreochromis niloticus exposed to lead, copper, and cadmium under temperature variations
Ragab G. Abdel Salam,, Samah M. Bassem, Eman Salah Abdel-Reheim, Mahmoud Abdel-Latif, Giulia Guerriero, Fagr Kh Abdel-Gawad,Isolation and partial characterization of serine proteases from jellyfish of the Antarctic region
Nataliia Raksha, Teyiana Halenova, Tetiana Vovk, Tetiana Beregova, Tetiana Maievska, Victor Tomchuk, Olexiy Savchuk, Ludmila OstapchenkoA computational statistical approach to assess cowpea [Vigna unguiculata (L.) Walp.] cultivars diversity and select elite genotypes from Agro-morphological and biochemical traits
Noel Dougba Dago, Jean Simon Konan Assouman, Saraka Didier Martial Yao, Nafan Diarrassouba