1. Home
  2. Volume 19, Number 1, February 2019
  3. Iqbalsyah

CULTIVATION CONDITIONS FOR PROTEASE PRODUCTION BY A THERMO-HALOSTABLE BACTERIAL ISOLATE PLS A

Teuku M. Iqbalsyah, Malahayati Malahayati, Atikah Atikah, Febriani Febriani

Abstract


Polyextremophiles have increasingly been utilised to produce thermostable enzymes with better stability in multiple extreme conditions. This study reports the screening results of four new bacterial isolates (PLS A, PLS 75, PLS 76 and PLS 80), isolated from an under water hot springs, in producing thermo-halostable protease enzyme. Optimum cultivation conditions for the protease production were also studied. Screening of protease-producing isolates was conducted using Thermus solid medium enriched with 3% skim milk and 0.5% casein. The growth of the isolates showing protease activity was monitored by measuring the cell dry weight and protease activity during 24 h cultivation period. The activity was also measured at various cultivation conditions, i.e. temperature, pH and salt concentrations. Amongst the four isolates, only PLS A showed the ability to produce protease. The optimum cultivation conditions for protease production were observed at 65°C, pH 7 for 18 h incubation. The activity increased with the addition of 1% NaCl concentration (0.085 Unit/mL). The ability of PLS A isolate to produce thermo-halostable protease was encouraging as they could potentially be used in industries requiring the enzyme with multiple extremes. 

Keywords


Thermophilic, halophilic, protease, thermozyme,

References


Bonch-Osmolovskaya, E.; Atomi, H. Editorial overview: Extremophiles: From extreme environments to highly stable biocatalysts. Curr. Opin. Microbiol. 2015, 25, vii-viii.

Kumar, L.; Awasthi, G.; Singh, B. Extremophiles: A novel source of industrially important enzymes. Biotechnol. 2011, 10 (2), 121-135.

Dalmaso, G.Z.L.; Ferreira, D.; Vermelho, A.B. Marine extremophiles: A source of hydrolases for biotechnological applications. Mar. Drugs. 2015, 13, 1925-1965.

Elleuche, S.; Schroder, C.; Sahmn, K.; Antranikian, G. Extremozymes-biocatalysts with unique properties from extremophilic microorganisms. Curr. Opin. Biotechnol. 2014, 29, 116-123.

de Lourdes Moreno, M.; Perez, D.; Garcia, M.T.; Mellado, E. Halophilic bacteria as a source of novel hydrolytic enzymes, Life, 2013, 3: 38-51.

Doukyu, N.; Ogino, H. organic solvent-tolerant enzyme, Biochem. Eng. J. 2010, 48, 270-282.

Kim, J.; Dordick, J.S. Usual salt and solvent dependent of a protease from an extreme halophiles. Biotech. Bioeng. 1997, 55, 471-479.

Gupta, R.; Beg. Q.K.; Lorenz, P. Bacterial alkaline proteases: molecular approaches and industrial applications. Appl. Microbiol. Biotechnol. 2002, 59, 15–32.

Anson, M. L., 1938. The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin, J. Gen. Physiol. 22, 79-89.

Encyclopedia Britannica, (2018, October 6), Chemical and physical properties of seawater: Dissolved inorganic substances, Retrieved from: https://www.britannica.com/science /seawater/Dissolved-inorganic-substances.

Lewis, E.L.; Perkin, R.G. Salinity: Its definition and calculation. J. Geophys. Res.1978, 83 (C1), 466-478.

Todar K. Todar's online textbook of bacteriology (2018, October 6), Retrieved from: http://textbookofbacteriology.net/ nutgro_3.html

Fuad, M. A.; Rahmawati, R.; Mubarik, N.R. Production and partial characterization of thermostable alkaline protease from Bacillus thermoglusidasius AF-01. Microbiol. Indonesia 2004, 9, 29-35.

Sharma, K.M.; Kumar, R.; Panwar, S.; Kumar, A. Microbial alkaline proteases: Optimization of production parameters and their properties. J. Gen. Eng. Biotechnol. 2017, 15, 115-126.

Chen, X.-G.; Stabnikova, O.; Tay, J.-H.; Wang, J.-Y.; Tay, S.T.-L. Thermoactive extracellular proteases of Geobacillus caldoproteolyticus, sp. nov., from sewage sludge. Extremophiles 2004, 8 (6), 489-498.

Asokan, S.; Jayanthi C. Alkaline protease production by Bacillus licheniformis and Bacillus coagulans. J. Cell Tissue Res., 2010, 10, 2119-2123.

Naidu, K.S.B., and Devi, K.L., Optimization of thermostable alkaline protease production from species of Bacillus using rice bran, African J. Biotechnol. 2005, 4(7), 724-726.

Chu, H.-W. Optimization of extracellular alkaline protease production from species of Bacillus. J. Ind. Microbiol. Biotechnol. 200734: 241-245.

Bkhairia, I.; Mhamdi, S.; Jridi, M.; Nasri, M. New acidic proteases from Liza aurata viscera: Characterization and application in gelatin production. Int. J. Biol. Macromol. 2016, 92, 533–542.

Ma, X.; Liu, Y.; Li, Q.; Liu, L.; Yi, L.; Ma, L.; Zhai, Z. Expression, purification and identification of a thermolysin-like protease, neutral protease I, from Aspergillus oryzae with the Pichia pastoris expression system. Prot. Exp. Purif. 2016, 128, 52-59.

Ibrahim, A.S.S, Salamah, A.A.A., Elbadawi, Y.B., El-Tayeb, M.A., Ibrahim, S.S.S. Production of extracellular alkaline protease by new halotolerant alkaliphilic Bacillus NPST-AK15 isolated from hyper saline soda lakes. Electronic J. Biotechnol. 2015,18, 236-243.

Yin, J.; Chen, J.C.; Wu, Q.; Chen, G.Q. Halophiles, coming stars for industrial biotechnology Biotechnol. Adv. 2015, 33, 1433–1442


Full Text: PDF

DOI: 10.24815/jn.v19i1.11971

Refbacks

  • There are currently no refbacks.