Production and characterization of ?-amylase from lactic acid bacteria isolated with whey

Year 2018, Volume: 11 Issue: 2, 115 - 122, 15.08.2018

İhsan Rezzukoğlu Sema Agüloğlu Fincan Barış Enez

Abstract

In this study, lactic acid bacteria were isolated from whey, to provide economic and biotechnological benefits. These bacteria which were isolated were described with the aid of morphological, physiological, and biochemical tests and named as PS-2A, PS-3B, PS-3C and PS-4B. And then, by investigating extracellular α–amylase production capabilities of these bacteria, PS-2A bacterium which showed the best activity was selected. It was found that bacterium PS-2A was round-like, Gram positive, catalase negative, active. For produce the PS-2A; the optimum incubation time as 16th hours, optimum pH as 7.0 and optimum incubation temperature was determined as 35°C. It was found that PS-2A conducted its optimum α–amylase activity at the 20th hour in LB medium performed, and optimum pH and temperature values of α–amylase activity were 7.0 and 35°C, respectively. In carbon sources; 1% soluble starch was increased the α–amylase production 2 fold. In nitrogen sources; it was determined that with 1% ammonium chlorid the α–amylase production was increased

Keywords

whey, lactic acid bacteria, -amylase, production, characterization

Peynir altı suyundan izole edilen laktik asit bakterisinden α -amilaz üretimi ve karakterizasyonu

Abstract

Bu çalışmada, peynir altı suyundan, ekonomik ve biyoteknolojik olarak fayda sağlamak amacıyla laktik asit bakterileri izole edildi. İzole edilen bakteriler morfolojik, fizyolojik ve biyokimyasal testler yardımıyla tanımlandı. Bakterilerin ekstraselüler α-amilaz üretme yeteneği araştırıldı. En iyi α -amilaz aktivitesi gösteren Lactococcus sp. PS2A ile çalışmalara devam edildi. Lactococcus sp. PS-2A bakterisinin kok yuvarlak şekilli, Gram pozitif, katalaz negatif, hareketli olduğu belirlendi. Bakteri üremesinin optimum koşulları pH 7.0, 35°C ve 16. saat olarak tespit edildi. Lactococcus sp. PS2A’nın en yüksek α-amilaz aktivitesini; LB besi yerinde, pH 7.0, 35°C ve 20. saatte gösterdiği belirlendi. Karbon kaynaklarında %1’lik çözünebilir nişastanın α-amilaz üretimini iki kat arttırdığı saptandı. Azot kaynaklarından %1’lik amonyum klorür ile α-amilaz üretiminin arttığı tespit edildi

Keywords

peynir altı suyu, laktik asit bakterisi, α-amilaz, üretim, karakterizasyon

References

• Adnan, A. F. M., Tan, İ. K. P. (2007). Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresource Technology, 98, 1380–1385.
• Agüloğlu, S., Enez, B. (2014). Purification, and characterization of α-amylase from thermophilic Geobacillus stearothermophilus. Starch/Stärke 66, 182–189
• Agüloğlu Fincan, S., Enez, B. Ozdemir, S., Matman Bekler, F. (2014). Purification and characterizasyon of thermostable α-amylase from thermophilic Anoxbacillus flavithermus. Carbohydrate Polymers, 102, 144-150.
• Ammor, M. S., Flo´rez, A. B., Mayo, B. (2007). Antibiotic resistance in non-enterococcal lactic acid bacteria and bifidobacteria. Food Microbiology, 24:559–570.
• Asgher, M., Asad, M. J., Rahma, S.U., Legge, R.L. (2007). A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. Journal of Food Engineering, 79, 950–955.
• Ayad, E.H.E., Verheul, A., Wouters, J.T.M., Smit, G. (2002). Antimicrobial-producing wild lactococci isolated from artisanal and non-dairy origins. International Dairy Journal, 12, 145–150.
• Badis, A., Guetarni, D., Boudjema, B. M., Henni, D.E., Kihal, M. (2004). Identification and technological properties oflactic acid bacteria isolated from raw goat milk of four Algerian races. Food Microbiology, 21,579–588.
• Bernfeld, P. (1955). Enzymes Carbohydrate Metabolism. In Methods In Enzymology Academic Press, 17,149-158.
• Burhan, A., Nisa, U., Gokhan, C., Omer, C., Ashabil, A., & Osman, G. (2003). Enzymatic properties of a novel thermophilic, alkaline and chelator resistant amylase from an alkalophilic Bacillus sp. isolate ANT-6. Process Biochemistry, 38, 1397–1403.
• Castro, P. M. L., Hayter, P. M., Ison, A. P., Bull, A. T. (1992). Application of statistical design to the optimization of culture medium for recombinant interferon-gamma production by Chinese hamster ovary cells. Applied Microbiology and Biotechnology, 38, 84–90.
• Çağırgan, H. (2004). Biotyping of Lactococcus garvieae isolated from Turkey. Ege University Journal of Fisheries & Aquatic Sciences, 21, 267– 269.
• Evans, J. J., Pasnik, D. J., Klesius, P. H., Al-Ablani, S. (2006). First report of Streptococcus agalactiae and Lactococcus garvieae from a wild bottlenose dolphin (Tursiops truncatus). Journal of Wildlife Diseases, 42, 561–569.
• Gupta, R., Gigras. P., Mohapatra. H., Goswami, V.K., Chauhan, B. (2003). Microbial α- amylase: biotechnological perspective. Enzyme and Microbial Technology, 38: 1599–1616.
• Jin, B., Van-Leeuwen, J. H., Patel, B. (1999). Mycelial morphology and fungal protein production from starch processing wastewater in submerged cultures of Aspergillus oryzae. Process Biochemistry, 34, 335–340.
• Kav, K., Erganiş, O. (2007). Konya bölgesinde bulunan gökkuşağı alabalığı (Oncorhynchus mykiss) çiftliklerinden Lactococcus garvieae izolasyonu, identifikasyonu ve fenotipik özelliklerinin belirlenmesi. Veterinerlik Bilimleri Dergisi, 23, 7–17.
• Kim, J., Sunako, M., Ono, H. Murooka, Y., Fukusaki, E., Yamashita, M. (2008). Characterization of gene encoding amylopullulanase from plant-originated lactic acid bacterium, Lactobacillus plantarum L137. Journal of Bioscience and Bioengineering, 106, 449–459.
• Konsula, Z., Liakopoulou-Kyriakides, M. (2004). Hydrolysis of starches by the action of an α-amylase from Bacillus subtilis. Process Biochemistry, 39, 1745–1749.
• Kubilay, A., Altun, S., Uluköy, G., Diler, Ö. (2005). Lactococcus garvieae suşlarının antimikrobiyal duyarlılıklarının belirlenmesi. Süleyman Demirel Üniversitesi Eğirdir Su Ürünleri Fakültesi Dergisi, 1, 39-48.
• amylase from a newly isolated Bacillus sp. YX-1: purification and
• characterization. Bioresource Technology, 99, 4315– 4320.
• Lowry, O.H., Rosebrough. N.J., Farr, A.L. (1951). Protein measurement with the folin phenol reagent, Journal of Biological Chemistry, 193, 265– 275.
• Marek, P., Nair, M. K. M., Hoagland, T., Venkitanarayanan, K. (2004). Survival and growth characteristics of Escherichia coli O157.H7 in pasteurized Cheddar cheese whey. International Journal of Food Microbiology, 94, 1–7.
• Ortakaya, V., Agüloğlu Fincan, S., Enez, B. (2017). α-Amylase from Bacillus sımplex- productıon,characterızation and partial purifıcation, Fresenius Environmental Bulletin, 26, 4446-4455.
• Ravelo, C., B. Magarinos, Romalde, J. L., Toranzo, A. E. (2001). Conventional versus miniaturized systems for the phenotypic characterization of Lactococcus garvieae strains. Bulletin European Association of Fish Pathologists, 21, 136–144.
• Sanni, A.I., Morlon-Guyot, J., Guyot, J.P. (2002). New efficient amylase-producing strains of Lactobacillus plantarum and L. fermentum isolated from different Nigerian traditional fermented foods. International Journal of Food Microbiology, 72, 53–62.
• Sengun, I. Y., Nielsen D. S., Karapinar M., Jakobsen M. (2009). Identification of lactic acid bacteria isolated fromarhana, a traditional Turkish fermented food. International Journal of Food Microbiology, 135, 105–111.
• Sharifiyazdi, H., Akhlaghi, M., Tabatabaei, M., Mostafavi Zadeh, S. M. (2010). Isolation and characterization of Lactococcus garvieae from diseased rainbow trout (Oncorhynchus mykiss,Walbaum) cultured in Iran. Iranian Journal of Veterinary Research, 11, (4), 33.
• Soltani, M., Nikbakht, G.H., Ebrahimzadeh, H. A., Ahmadzadeh, N. (2008). Epizootic outbreaks of Lactococcus garvieae in farmed rainbow trout (Oncorhynchus mykiss) in Iran. Bulletin European Association of Fish Pathologists, 28, 209–214.
• Souza, B.M. S., Borgonovi, T.F., Casarotti, S. N., Todorov ,S. D., Penna, A.L.B. (2018). Lactobacillus casei and Lactobacillus fermentum Strains Isolated from Mozzarella Cheese: Probiotic Potential, Safety, Acidifying Kinetic Parameters and Viability under Gastrointestinal Tract Conditions. Probiotics and Antimicrobial Proteins, https://doi.org/10.1007/s12602-018-9406-y.
• Vendrell, D., Balcázar, J. L., Ruiz-Zarzuela, I., de Blas, I., Gironés, O., Mşzquiz, J. L. (2006). Lactococcus garvieae in fish: A review. Comparative Immunology, Microbiology and Infectious Diseases, 29, 177-198.
• Yerlikaya, O., Kınık, Ö., Akbulut, N. (2010). Peyniraltı Suyunun Fonksiyonel Özellikleri ve Peyniraltı Suyu Kullanılarak Üretilen Yeni Nesil Süt Ürünleri. GIDA, 35, 289–296.
• Yücel Şengün, İ. (2011). Lactic acid bacteria used in the production of fermented foods. Biological Diversity andConservation 4, 42– 53
• Zamfir, M., Vancanneyt, M., Makras, L., Vaningelgem, F., Lefebvre, K., Pot, B., Swings, J., De Vuyst, L. (2006). Biodiversity of lactic acid bacteria in Romanian dairy products. Systematic and Applied Microbiology, 29, 487–495..