Sains Malaysiana 47(1)(2018): 91–97
http://dx.doi.org/10.17576/jsm-2018-4701-11
Characterization and
Screening of Lipolytic Bacteria from Thai Fermented Fish
(Pencirian dan Penyaringan Bakteria Lipolitik daripada Ikan
Pekasam Thai)
RUNGSIMA DAROONPUNT1, NAOTO TANAKA2, MASATAKA UCHINO3 & SOMBOON TANASUPAWAT1*
1Department
of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn
University, Bangkok 10330, Thailand
2NODAI
Culture Collection Center (NRIC), Faculty of Applied Bio-Science, Tokyo
University of Agriculture, Tokyo 156-8502, Japan
3Department
of Applied Biology and Technology, Faculty of Applied Bio-science, Tokyo, University
of Agriculture, Tokyo 156-8502, Japan
Diserahkan:
10 Ogos 2015/Diterima: 6 Julai 2017
ABSTRACT
Nine bacterial strains were isolated from Thai
fermented fish by the standard dilution technique using JCM no.
377 medium. The rod shaped isolate FN2-3 was identified as Virgibacillus dokdonensis while isolates FN2-3
and FN6-6 as V. halodenitrificans, FN1-13
as Corynebacterium variabile, FN1-10 as Oceanobacillus
iheyensis and FN3-7 was Bacillus
amyloliquefaciens subsp. plantarum. The coccal isolates, FN6-1
and FN6-7 were Staphylococcus saprophyticus subsp. bovis and FN6-8 was S. saprophyticus subsp. saprophyticus,
based on their phenotypic characteristics and 16S rRNA gene sequence analyses
at a 99.63-100% sequence similarity. Their lipase activity in complex medium (CM), CM medium with 1% (v/v) Tween 20 or CM medium
with 1% (v/v) Tween 80 ranged from 1.12 ± 0.03-3.77 ± 0.04 unit/mL, with the
highest lipase activity found with V. dokdonensis FN1-8
cultivated with CM with 1% (v/v) Tween 80.
Keywords: Fermented fish; halophilic bacteria; lipase; lipolytic
bacteria
ABSTRAK
Sembilan strain bakteria telah dipencilkan daripada
ikan pekasam Thai melalui teknik kecairan piawai yang menggunakan
medium JCM no. 377. Pencilan berbentuk rod FN2-3
dikenal pasti sebagai Virgibacillus
dokdonensis manakala pencilan FN2-3 dan FN6-6
sebagai V. Halodenitrificans, FN1-13 sebagai Corynebacterium
variabile, FN1-10 sebagai Oceanobacillus iheyensis dan FN3-7
sebagai Bacillus amyloliquefaciens subsp. plantarum.
Pencilan kokus FN6-1
dan FN6-7 adalah Staphylococcus saprophyticus subsp.
bovis dan FN6-8 S. saprophyticus subsp. saprophyticus,
berdasarkan ciri fenotip dan 16S rRNA analisis jujukan gen pada
satu persamaan turutan 99.63-100%. Aktiviti lipase mereka dalam
medium kompleks (CM),
medium CM 1% (v/v) Tween 20 atau CM medium
dengan 1% (v/v) Tween 80 berjulat dari 1.12 ± 0.03-3.77 ±
0.04 unit/mL dengan aktiviti lipase tertinggi dilihat pada V.
dokdonensis FN1-8
yang ditanam dengan 1% (v/v) Tween 80 CM.
Kata kunci: Bakteria
halofili lipase; bakteria lipolitik; ikan pekasam
RUJUKAN
Barrow, G.I. & Feltham, R.K.A.
1993. Cowan and Steel’s Manual for the Identification of Medical Bacteria.
3rd ed. Cambridge: Cambridge University Press. pp. 331.
Boutaiba, S., Bhatnagar, T., Hacene,
H., Mitchell, D.A. & Baratti, J.C. 2006. Preliminary characterization of a
lipolytic activity from an extremely halophilic archaeon, Natronococcus sp. Journal of Molecular Catalysis B: Enzymatic 41: 21-26.
Camacho, R.M., Mateos, J.C.,
González-Reynoso, O., Prado, L.A. & Córdova, J. 2009. Production and
characterization of esterase and lipase from Haloarcula marismortui. Journal
of Industrial Microbiology and Biotechnology 36: 901-909.
Chun, J., Lee, J., Jung, Y., Kim, M.,
Kim, S., Kim, B.J. & Lim, Y. 2007. EzTaxon: A web based tool for the
identification of prokaryotes based on 16 ribosomal RNA gene sequences. International
Journal of Systematic and Evolutionary Microbiology 57: 2259-2261.
Collins, M.D. 1987. Transfer of Arthrobacter
variabilis (Muller) to the Genus Corynebacterium, as Corynebacterium
variabilis comb. nov. International Journal of Symstematic Bacteriology 37:
287-288.
Felsenstein, J. 1985. Confidence limits
on phylogenies: An approach using the bootstrap. Evolution 39: 783-791.
Fukami, K., Ishiyama, S., Yaguramaki,
H., Masuzawa, T., Nabeta, Y., Endo, K. & Shimoda, M. 2002. Identification
of distinctive volatile compounds in fish sauce. Journal of Agricultural and
Food Chemistry 50: 5412-5416.
García-Lepe, R., Nuero, O.M., Reyes, F.
& Sanatamaría, F. 1997. Lipases in autolysed cultures of filamentous fungi. Letters in Applied Microbiology 25: 127-130.
Gayathri, V.R., Perumal, P., Mathew,
L.P. & Prakash, B. 2013. Screening and molecular characterization of
extracellular lipase producing Bacillus species from coconut oil mill
soil. International Journal of Science and Technology 2: 502-509.
Ghasemi, Y., Rasoul-Amini, S., Kazemi,
A., Zarrini, G., Morowvat, M.H. & Kargar, M. 2011. Isolation and
characterization of some moderately halophilic bacteria with lipase activity. Microbiology 80: 483-487.
Gupta, R., Gupta, N. & Rathi, P.
2004. Bacterial lipases: An overview of production, purification and
biochemical properties. Applied Microbiology and Biotechnology 64:
763-781.
Hajek, V., Meugnier, H., Bes, M., Brun,
Y., Fiedler, F., Chmela, Z., Lasne, Y., Fleurette, J. & Freney, J. 1996. Staphylococcus
saprophyticus subsp. bovis subsp. nov., isolated from bovine
nostrils. International Journal of Systematic Bacteriology 46: 792-796.
Hasan, F., Shah, A.A. & Hameed, A.
2006. Industrial applications of microbial lipases. Enzyme and Microbial
Technology 39: 235-251.
Higaki, S. & Morohashi, M. 2003. Propionibacterium
acnes lipase in seborrheic dermatitis and other skin diseases and Unsei-in. Drugs under Experimental and Clinical Research 29: 157-159.
Kumar, D., Kumar, L., Nagar, S., Raina,
C., Parshad, R. & Gupta, V.K. 2012. Screening, isolation and production of
lipase/ esterase producing Bacillus sp. strain DVL2 and its potential
evaluation in esterification and resolution reactions. Archives of Applied
Science Research 4: 1763-1770.
Leifson, E. 1963. Determination of
carbohydrate metabolism of marine bacteria. Journal of Bacteriology 85:
1183-1184.
Li, H. & Zhang, X. 2005.
Characterization of thermostable lipase from Thermophilic Geobacillus sp.
TW1. Protein Expression and Purification 42: 153-159.
Li, X., Qian, P., Wu, S.G. & Yu,
H.Y. 2014. Characterization of an organic solvent-tolerant lipase from Idiomarina sp. W33 and its application for biodiesel production using Jatropha oil. Extremophiles 18: 171-178.
Lu, J., Nogi, Y. & Takami, H. 2001. Oceanobacillus iheyensis gen. nov., sp. nov., a deep-sea extremely
halotolerant and alkaliphilic species isolated from a depth of 1050 m on the
Iheya Ridge. FEMS Microbiology Letters 205: 291-297.
Martin, S., Marquez, M.C.,
Sanchez-Porro, C., Mellado, E., Arahal, D.R. & Ventosa, A. 2003. Marinobacter
lipolyticus sp. nov., a novel moderate halophile with lipolytic activity. International
Journal of Systematic and Evolutionary Microbiology 53: 1383-1387.
Miettinen, H., Nyyssölä, A., Rokka, S.,
Kontkanen, H. & Kruus, K. 2013. Screening of microbes for lipases specific
for saturated medium and long-chain fatty acids of milk fat. International
Dairy Journal 32: 61-67.
Mrozik, A., Hubert-Kocurek, K. &
Łabużek, S. 2006. Lipases of genera Pseudomonas and Burkholderia and their applications in biotechnology. Postępy Mikrobiologii 45:
19-26.
Niazi, A., Manzoor, S., Bejai, S.,
Meijer, J. & Bongcam-Rudloff, E. 2014. Complete genome sequence of a plant
associated bacterium Bacillus amyloliquefaciens subsp. plantarum UCMB5033. Standards in Genome Sciences 9: 718-725.
Namwong, S., Tanasupawat, S., Smitinont, T., Visessanguan,
W., Kudo, T. & Itoh, T. 2005. Isolation of Lentibacillus juripiscarius sp. nov. from fish sauce in Thailand. International Journal of Systematic
and Evolutionary Microbiology 55: 315-320.
Noureddini,
H., Gao, X. & Philkana, R.S. 2005. Immobilized Pseudomonas cepacia lipase
for biodiesel fuel production from soybean oil. Bioresource Technology 96:
769-777.
Ruiz,
C., Javier Pastor, F. & Diaz, P. 2003. Isolation and characterization of Bacillus sp. BP-6 LipA, a Ubiquitous lipase among mesophilic Bacillus species. Letters in Applied Microbiology 37: 354-359.
Saitou,
N. & Nei, M. 1987. The neighbor-joining method: A new method for
reconstructing phylogenetic trees. Molecular Biology and Evolution 4:
406-425.
Sharma,
R., Christi, Y. & Banerjee, U.C. 2001. Production, purification,
characterization and application of lipases. Biotechnology Advances 19:
627-662.
Sirisha,
E., Rajasekar, R. & Lakshmi Narasu, M. 2010. Isolation and optimization of
lipase producing bacteria from oil contaminated soils. Advances in
Biological Research 4: 249-252.
Snellman,
E.A., Sullivan, E.R. & Colwell, R.R. 2002. Purification and properties of
the extracellular lipase, LipA, of Acinetobacter sp. RAG-1. Federation
of European Biochemical Societies 269: 5771-5779.
Tamura,
K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. 2011.
MEGA5: Molecular evolutionary genetics analysis using maximum likelihood,
evolutionary distance, and maximum parsimony methods. Molecular Biology and
Evolution 28: 2731-2739.
Tanasupawat,
S. & Komagata, K. 2001. Lactic acid bacteria in fermented foods in
Southeast Asia, in Microbial Diversity in Asia: Technology and Prospects, edited
by B.H. Nga, H.M. Tan, and K. Suzuki). Singapore: World Scientific Publishing
Co. Pte. Ltd. pp. 43-59.
Walavalkar,
G.S. & Bapat, M.M. 2001. Staphylococcus warneri BW 94 - A new source
of lipase. Indian Journal of Experimental Biology 40: 1280-1284.
Yamada,
Y., Katsura, K., Kawasaki, K., Widyastuti, Y., Saono, S., Seki, T., Uchimura,
T. & Komagata, K. 2000. Asaia bogorensis gen. nov., sp. nov., an
unusual acetic acid bacterium in the α-Proteobacteria. International
Journal of Systematic and Evolutionary Microbiology 50: 823-829.
Yoon,
J.H., Kang, S.J., Lee, S.Y., Lee, M.H. & Oh, T.K. 2005. Virgibacillus
dokdonensis sp. nov., isolated from a Korean island, Dokdo, located at the
edge of the East Sea in Korea. International Journal of Systematic and
Evolutionary Microbiology 55: 1833-1837.
Yoon,
J.H., Oh, T.K. & Park, Y.H. 2004. Transfer of Bacillus halodenitrificans Denariaz et al. 1989 to the genus Virgibacillus as Virgibacillus
halodenitrificans comb. nov. International Journal of Systematic and
Evolutionary Microbiology 54: 2163-2167.
*Pengarang
untuk surat-menyurat; email: Somboon.T@chula.ac.th
|