Citation: Suribabu K, Lalitha Govardhan T, Hemalatha KPJ (2014) Strain Improvement of Brevibacillus borostelensis R1 for Optimization of α-Amylase Production by Mutagens. J Microb Biochem Technol 6: 123-127. doi:10.4172/1948-5948.1000132 Copyright: © 2014 Suribabu K, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited AbstractPhysical and chemical mutagens are promising and are used for screening of high yielding strains. The exponential increase in the application of amylases in various fields has placed stress and demand in both qualitative improvement and quantitative enhancement through strain improvement. Ultraviolet light exerts its mutagenic effect by exciting electrons in molecules. The potent UV mutants which showed more than 20 mm zone of starch hydrolysis were screened and selected at 42% of survival time at 80minutes of exposure. The wild strain with fixed parameters yielded (3000 U/ml). The major findings of the strain improvement were out of ten mutants isolated, two (UV-3 and UV-10) showed 3000-4000 U/ml of amylase activity. The % of survival of Brevibacillus borstelensis R1 in Pikovskaya's medium was 25.75% at 120 minutes of exposure. Ten mutants (HNO 2 -10, HNO 2 -30, EMS-4, EtBr-40, EtBr-50, Acr-1, Acr-20, Acr-30, Acr-4 and 5′-FU-50) out of fifty mutants isolated showed 3000-4300U/ml of amylase activity, which was higher than the wild strain. The potent Bacillus species screened from marine water was Brevibacillus borstelensis R1. The α-amylase was found to be useful in bakery, food, fodder for poultry, automation dishwashing and laundry industries.
Bacteria have been regarded as treasure of many useful enzymes viz., amylases, proteases, lipases, hydrolases and reductases. Among them amylolytic enzymes have great biotechnological applications and economic exploitations. The production of α-amylases by fermentation had been thoroughly investigated and shown to be affected by a variety of physicochemical factors, such as the composition of the growth medium, the type of strain, cell growth, methods of cultivation, inoculum concentration, time of incubation, pH, temperature, salinity, carbon, nitrogen and mineral sources. The present study was carried out to optimize the α-amylase production of Brevibacillus borstelensis R1 using ten different media viz., Nutrient broth, Luria Bertain broth, Clarks & Lub medium, Pikovskaya's medium, Tendler's non-synthetic medium, Amylase production medium, Soluble starch beef extract medium, Soybean casein digest medium, Yeast extract peptone dextrose glucose medium and Tryptone glucose beef extract medium. Among these ten media, Pikovskaya's (PK) medium proved to be optimal for α-amylase production (1861±17U/ml). The optimized α-amylase production in PK medium by submerged fermentation (SmF) was subjected to varying physical parameters such as 24hrs incubation time, 2% inoculum size, 37 0 C, pH 7.0 and 1% NaCl. Alpha-amylase produced by B.borostelensis R1 have many applications in starch processing, desizing of textiles, paper sizing, detergent additive, bread improvement, ethanol production, sewage treatment, effluent treatment and other fermentation processes.
Numerous marine microorganisms secrete enzymes which can provide new insights and understanding of enzymes. Marine microorganisms have been attracting more attention as source for novel enzymes. Secondary screening is strictly essential in any systematic screening programme which helps in detection of useful bacteria in fermentation processes. Secondary screening also provides information pertaining to the effect of different components of the medium. This is valuable in designing the medium that may be attractive as far as economic consideration is concerned. Natural carbon source, Saccharum officinarum (5%) produced maximum α-amylase while Triticum vulgare (4%) produced very low α-amylase. Synthetic carbon source when supplemented with maltose (1%) and sucrose (4%) regulated higher production of amylase. Starch (2%), dextrose (3%) and galactose (4%) exhibited average effect on production whereas lactose (3%) and mannitol (3%) decreased production. The α-amylase was found to have many applications in the field of starch processing, textile industry, improving shelf life of bread, ethanol production, sewage treatment and effluent treatment.
Thermostability is a desired characteristic of most of the industrial enzymes. The production of α-amylases has been thoroughly investigated and observed that it was affected by a variety of physiochemical factors, such as the composition of the growth medium under various temperature parameters with constant pH 7.0. The enzyme production was assayed in submerged fermentation (SmF) condition. The optimum production of α-amylase by SmF was observed at 4 0 C in Clarks and Lub Medium (2551 ± 1 U/ml), Soybean Casein Digest Medium (920± 0.13 U/ml) and Tryptone Glucose Beef Extract Medium (260 U/ml). The optimum production of αamylase by SmF was observed at 25 0 C in Nutrient Broth (270 U/ml). The optimum production of α-amylase by SmF was observed at 37 0 C in Pikovskaya's Medium (3371 ± 0.50 U/ml), Tendler's Non-synthetic Medium (761 ± 0.50 U/ml), Amylase Production Medium (580 U/ml) and Soluble Starch Beef Extract Medium (1930 U/ml). The optimum production of α-amylase by SmF was observed at 50 0 C in Luria Bertain Broth (240 ± 0.12 U/ml) and Yeast extract peptone Glycerol Glucose Medium (265 U/ml). Thermal variability of α-amylase produced by Brevibacillus borostelensis R1 in various media may have a significant role in many applications like bakery industry, treatment of effluents from sago and rice industry, sewage water treatment, fodder production, laundry industry and textile industry.
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