Comparison on Effects of Temperature on Different Strains of Phytase Producing Bacteria Isolated from Malaysia’s Hot Spring

Anis Adilah Mustafa, N. A. Zulkifly, T. Abdullah, N. Alias
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Abstract

The main purpose of this research was to find the best growth curve for bacterial growth and the optimum temperature for the production of phytase from different potential phytase producing bacterial strains. A total of four strains used were originally isolated from hot springs in Malaysia, which were in Labis, Johor (L3), Dusun Tua, Selangor (RT), Ulu Legong, Kedah (A) and Ranau, Sabah (B9). Nutrient Agar (NA) and modified Phytase Screening Medium (PSM) liquid media were used for the culture enrichment while optimisation was carried out through batch culture method using a shake-flask scale. Strains growth and enzyme activity were quantitatively measured at different temperatures at (30°C and 37°C) values. Enzyme activity was determined according to the reaction of the phytase with its substrate (sodium phytate) and expressed in units of phytase activity (U/ mL). As for the overall, strain L3 (from Labis, Johor) exhibit promising rate of Pi released in the media at 30°C and 37°C, with optimum phytase activity values of 0.2047 U/mL and 0.2195 U/mL, respectively. The pH of the cultures was also measured, where it shows that strains grown in cultures at 37°C produced a higher phytase activity and resulting a lower reading of pH compared when grown at 30°C. All around, L3 strains has the lowest value of pH when cultured at 30°C and 37°C, with the pH value of 3.62 and 2.37, respectively.  From the result obtained, the lower pH indicates the process of phytic acid degradation take place by the phytase in producing inorganic phosphate (Pi) due to the accumulation of organic acid. Since these bacterial strains were originally taken from Hot Springs, further analysis of temperature optimization using 55°C and even 60°C should be carried out. In the future, biochemical research and molecular identification may also be carried out to identify molecular identity in the strains.
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温度对马来西亚温泉不同产植酸酶菌株影响的比较
本研究的主要目的是寻找不同产植酸酶潜力菌株的最佳生长曲线和产植酸酶的最佳温度。所使用的四种菌株最初是从马来西亚的温泉中分离出来的,分别是柔佛州拉比斯(L3)、雪兰莪州Dusun Tua (RT)、吉打州乌鲁勒贡(A)和沙巴拉瑙(B9)。采用营养琼脂(NA)和改良植酸酶筛选培养基(PSM)液体培养基进行培养富集,并采用摇瓶规模分批培养法进行优化。在不同温度下(30°C和37°C)定量测定菌株生长和酶活性。根据植酸酶与其底物(植酸钠)的反应测定酶活性,以植酸酶活性单位(U/ mL)表示。总体而言,菌株L3(来自柔佛Labis, Johor)在30°C和37°C的培养基中表现出良好的Pi释放率,最佳植酸酶活性值分别为0.2047 U/mL和0.2195 U/mL。还测量了培养物的pH值,结果表明,与在30°C培养的菌株相比,在37°C培养的菌株产生更高的植酸酶活性,pH值读数较低。从各方面来看,L3菌株在30℃和37℃培养时pH值最低,分别为3.62和2.37。从得到的结果来看,较低的pH值表明由于有机酸的积累,植酸酶在产生无机磷酸盐(Pi)时发生了植酸降解过程。由于这些菌株最初是从温泉中提取的,因此需要在55°C甚至60°C下进行进一步的温度优化分析。今后还可开展生化研究和分子鉴定,对菌株进行分子鉴定。
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