Beta Galactosidase Enzyme Activities of Lactobacillus and Bifidobacterium Genus (Turkish with English Abstract)

Yıl 2014, Cilt: 39 Sayı: 4, 211 - 218, 01.08.2014

Yasemin Kılıç Zehra Nur Yüksekdağ Hazer Yüksekdağ

https://izlik.org/JA55WB56YF

Öz

In this study total 42 bacteria were used; 39 Lactobacilllus originated from human, food and animal and 3 Bifidobacterium isolated from new born baby feces. The β-galactosidase enzyme activities and specific activities were determined by using o-nitrophenyl-b-D-galactopyranoside (o-NPG) as a substrate. The highest specific enzyme activities observed among Lactobacilli cultures were at L. fermentum ZYN17 (2.468 U/mg), L. casei LB65 (1.116 U/mg), L. rhamnosus GD11 (1.034 U/mg), and L. acidophilus BAZ36 (0.947 U/mg) strains and the highest enzyme activities observed among Bifidobacterium was at B. breve A26 (0.726 U/mg) strain. The enzyme activities of bacteria were also determined qualitatively by using the 5-bromo-4-chloro-3-indolyl-β-D-galaktoside (X-gal) substrate compound. The enzyme optimization of ZYN17 strain with highest specific β-galactosidase activity was achieved. The optimum pH was determined as 6.8, optimum temperature was 37 ºC and the optimum buffer was potassium phosphate buffer for the enzyme.

Anahtar Kelimeler

Lactobacilllus , Bifidobacterium , β-galactosidase enzyme activity

Lactobacillus ve Bifidobacterium Cinsi Bakterilerin Beta Galaktosidaz Enzim Aktivitelerinin Belirlenmesi

https://izlik.org/JA55WB56YF

Öz

Bu çalışmada, insan, gıda ve hayvan kaynaklı 39 Lactobacilllus cinsine ait ve yeni doğan gaitasından izole edilmiş 3 Bifidobacterium cinsine ait toplam 42 bakteri kullanılmıştır. O-nitrofenil-beta-D-galaktosit (o-NPG) substrat olarak kullanılarak, kültürlerin β-galaktosidaz enzim ve spesifik aktiviteleri belirlenmiştir. Lactobacillus cinsine ait kültürlerden L. fermentum ZYN17 (2.468 U/mg), L. casei LB65 (1.116 U/mg), L. rhamnosus GD11 (1.034 U/mg) ve L. acidophilus BAZ36 (0.947 U/mg) suşlarının, Bifidobacterium cinsine ait kültürlerden de B. breve A26 (0.726 U/mg) suşunun en yüksek spesifik aktivite yeteneğine sahip oldukları tespit edilmiştir. Ayrıca, bakterilerin 5-brom-4-klor-3-indolil-β-D-galaktopiranosit (X-gal) substrat bileşiğiyle de nitel olarak enzim aktivitesinin varlığı değerlendirilmiştir. Yüksek spesifik β-galaktozidaz aktivitesi gösteren ZYN17 suşuna ait β-galaktozidaz enziminin optimizasyonu yapılmıştır. β-galaktozidaz enziminin optimum pH’sı 6.8, optimum sıcaklığı 37 ºC ve optimum tamponun potasyum fosfat tamponu olduğu belirlenmiştir.

Anahtar Kelimeler

Lactobacilllus , Bifidobacterium , β-galaktosidaz enzim aktivitesi  

Kaynakça

• Gheytanchi E, Heshmati F, Shargh KB, Nowroozi J, Movahedzadeh F. 2010. Study on E-galactosidase enzyme produced by isolated lactobacilli from milk and cheese. Afr J Microbiol Res, 4, 454-458.
• Karasova P, Spiwok V, Mala S, Kralova B, Russell NJ. 2002. Beta-galactosidase activity in psychrophic microorganisms and their potential use in food industry. Czech J Food Sci, 20(2), 43-47.
• Verma LM, Barrow JC, Kennedy JF, Puri M. 2012. Immobilization of E-d-galactosidase from Kluyveromyces lactison functionalized silicon dioxide nanoparticles: Characterization and lactose hydrolysis. Int J Biol Macromol, 50(2), 432-437.
• Panesar R, Panesar PS, Singh RS, Kennedy JF. 2011. Hydrolysis of milk lactose in a packed bed reactor system using immobilized yeast cells. J Chem Technol Biotechnol, 86(1), 42-46.
• Vonk RJ, Reckman GAR, Harmsen HJM, Priebe MG. 2012. Probiotics and Lactose Intolerance. Chapter 7. InTech Open, 7, 149-160.
• Singh VP, Sharma J, Babu S, Rizwanulla, Singla A. 2013. Role of probiotics in health and disease: a review. J Pak Med Assoc, 63(2), 253-257.
• Setty-Shah N, Maranda L, Candela N, Fong J, Dahod I, Rogol AD, Nwosu BU. 2013. Lactose Intolerance: Lack of Evidence for Short Stature or Vitamin D Deficiency in Prepubertal Children. PLOS ONE, 8, 1-9.
• Bayhan A, Yentür G. 1993. Laktoz ‹ntoleransı. GIDA, 18 (6), 385-388.
• Rolfe RD. 2000. The role of probiotic cultures in the control of gastrointestinal health. J Nutr, (Supplement).130 (2), 396-402.
• Sanders ME. 1999. Probiotics. Food Technol, 53 (11), 67-77.
• De Vrese M, Stegelmann A, Richter B, Fenselau S, Laue C, Schrezenmeir J. 2001. Probiotics- compensation for lactase insufficiency. Am J Clin Nutr, 73 (2), 421-429.
• Zhang W, Wang C, Huang CY, Yu Q, Liu HC, Zhang CW, Pei XF, Xu X, Wang GQ. 2012. Analysis of E-galactosidase production and their genes of two strains of Lactobacillus bulgaricus. Biotechnol Lett, 34 (6), 1067-1071.
• Shah NP, Otieno DO. 2007. Endogenous E-glucosidase and E-galactosidase activities from selected probiotic microorganisms and their role in isoflavone biotransformation in soymilk. J Appl Microbiol, 103 (4), 910-917.
• Kara F. 2004. Release and characterization of beta-galactosidase from Lactobacillus plantarum. M.C. Thesis, Department of Biotechnology, Middle East Technical University, 89p.
• Ismail SAA, El-Mohamady Y, Helmy WA, Abou-Romia R, Hashem AM. 2010. Cultural condition affecting the growth and production of E-galactosidase by Lactobacillus acidophilus NRRL 4495. Aust J Basic Appl Sci, 4 (10), 5051-5058.
• Citti JE, Sandine WE, Elliker PR. 1965. E-Galactosidase of Streptococcus lactis. J Bacteriol, 89 (4), 937-942.
• Li J, Zhang W, Wang C, Yu Q, Dai R, Pei X. 2012. Lactococcus lactis expressing food-grade E-galactosidase alleviates lactose intolerance symptoms in post-weaning Balb/c mice. Appl Microbiol Biotechnol, 96 (6), 1499-506.
• Jokar A, Karbassi A. 2011. In-house production of lactose-hydrolysed milk by beta-galactosidase from Lactobacillus bulgaricus. J Agr Sci Technol, 13 (4), 577-584.
• Almeida CC, Lorena SLS, Pavan CR, Akasaka HMI, Mesquita MA. 2012. Beneficial effects on long-term consumption of a probiotic combination of Lactobacillus casei Shirota and Bifidobacterium breveYakult may persist after suspension of therapy in lactose- intolerant patients. Nutr Clin Pract, 27 (2), 247-251.
• Ouwehand AC, Salminen S, Isolauri E. 2002. Probiotics: An overview of beneficial effects. Antonie Leeuwenhoek, 82 (1-4), 279-289.
• Tari C, Ustok FI, Harsa S. 2010. Production of food grade E-galactosidase from artisanal yogurt strains. Food Biotechnol, 24 (1), 78-94.
• Mozumder NHMR, Akhtaruzzaman M, Bakr MA, Zohra FT. 2012. Study on isolation and partial purification of lactase (E-Galactosidase) enzyme from Lactobacillus bacteria isolated from yogurt. J Sci Res, 4 (1), 239-249.
• Meira MMS, Helfer EV, Velho VR, Lopes CF, Brandelli A. 2012. Probiotic potential of Lactobacillus spp. isolated from Brazilian regional ovine cheese. J Dairy Res, 79 (1), 119-127.
• Hsu CA, Yu RC, Chou CC. 2005. Production of E-galactosidase by Bifidobacteria as influenced by various culture conditions. Int J Food Microbiol, 104 (2), 197-206.
• Toba T, Tomita Y, Itoh T, Adachi S. 1981. E-Galactosidases of lactic acid bacteria: characterization by oligosaccharides formed during hydrolysis of lactose. J Dairy Sci, 64 (2), 185-192.
• Üstok FI. 2007. Production of E-Galactosidase Using Lactic Acid Bacteria and Optimisation of Fermentation Parameters. (M.Sc), Fen Bilimleri Enstitüsü, ‹zmir ‹leri teknoloji Enstitüsü, ‹zmir, 35s.
• Iqbal S. 2011. Characterization of E-Galactosidase and Enzymatic Synthesis of Prebiotic Galacto-oligosaccharides. M.Sc. Hons., University of Natural Resources and Life Sciences, Vienna, 70 p.
• Kim JW, Rajagopal SN. 2000. Isolation and characterization of E-galactosidase from Lactobacillus crispatus. Folia Microbiol, 45, 29-34. 29. Pamuk F. 2011. Biyokimya, Gazi Kitabevi, Ankara, Türkiye, 272s.
• Tzortzis G, Goulas AK, Gibson GR. 2005. Synthesis of prebiotic galactooligosaccharides using whole cells of a noval strain Bifidobacterium bifidumNCIMB 41171. Appl Microbiol Biotechnol, 68 (3), 412-416.
• Rabiu BA, Jay AJ, Gibson GR, Rastall RA. 2001. Synthesis and fermentation properties of novel galacto-oligosaccharides by E-galactosidase from Bifidobacterium species. Appl Environ Microbiol, 67 (6), 2526-2530.