Purification, characterization and potential detergent industry application of a thermostable ?-amylase from Bacillus licheniformis RA31

?-Amylase is an endoamylase catalysing the degradation of starch into maltose, maltotriose and glucose. The enzyme isolated from microbial sources possess unique properties of thermostability thus making it a useful tool in the detergent industry. Here, we explored a strain of thermophilic bacteria Bacillus licheniformis for its potential application in detergent industry. The B. licheniformis RA31 was isolated from soil samples of hot spring in Rampur district of Himachal Pradesh, India and grown on optimized media to produce thermostable ?-amylase. The enzyme was ethanol precipitated, purified (12.93 fold, 55.52% yield and 621.93 U/mg specific activity) to homogeneity with a single band on SDS-PAGE (66 kDa) and native-PAGE (68 kDa). Purified enzyme displayed best activity in pH 8 buffer and ?80% activity was retained in pH 7 and 10. It showed temperature optima at 70°C. Its activity was decreased at 70°C (70% after 4 h), 80°C (65% after 4 h) and 90°C (50% after 1 h). The enzyme was stimulated (126%; 5 mM) by barium chloride. It was relatively stable in the presence of commercial detergents (109-125%), SDS (84%), Tween 20 (88%), EDTA (72%) and ?-ME (70% at 10 mM). Km and Vmax for the enzymatic hydrolysis of starch were 0.339 mg/mL and 1.450 mg/min, respectively. The enzyme revealed the highest specificity towards wheat starch granule (140% after 1 h) and SEM analysis displayed its biodegradation (2-10 h). Improved cleaning efficiency of potato curry stained fine cotton clothes were observed with enzyme assisted detergent advance treatment (0.02% w/v). The enzyme showed potential applications in detergent industry.

Isolation and molecular identification of α-amylase producing bacteria from hot water spring by 16S rRNA gene sequencing

Among different types of microbial enzymes, amylases are the most widely used in industries as they are produced in large quantity and in an economic way as compared to plants and animals. Moreover, thermostable amylase has significance as compared to the amylase from mesophiles. Therefore, hot water springs are explored to dig into its bacterial diversity. In the current study, we tried to isolate amylase producing bacteria from the soil and water samples collected from the hot water spring in Rampur, Himachal Pradesh, India. The samples were serially diluted before plating on the Luria Bertani agar plates. A total of 42 bacterial morphotypes were isolated and were screened for amylolytic activity by starch agar plate method. Among the 42 bacterial isolates 25 showed amylolytic activity. Production of amylase was carried out at different temperatures and pH to optimize the temperature and pH conditions for each isolate. All the 25 isolates were characterized based on morphology, biochemical tests and molecular analysis. Sequencing of 16S rRNA gene for the 25 isolates followed by BLAST search revealed a majority of them (19) identified as Bacillus licheniformis. Other isolates were identified as B. subtilis, B. safensis, B. halodurans, B. stratosphericus, Caldimonas hydrothermale and Exiguobacterium mexicanum. An attempt was made to amplify amyN gene which codes for α-amylase but successful amplification was achieved only from Bacillus licheniformis alone.

Emerging of RNA Viruses: A Threat of Epidemics Around-The-Clock.

Viral infections are global public health concern and RNA viruses are the major cause of morbidity and mortality across the world due to their high error rate of replication and better adaptability inside the host cell. Some of the recent viral outbreaks around the globe are mainly hepatitis and its subtypes, influenza and its subtypes, Japanese encephalitis, dengue, ebola and the chikungunya. Vaccines are available only for some of these diseases. Therefore, organisation comprising WHO in accordance with the International Health Regulations of 2005 keeps on to track the evolving infectious diseases and the Global Outbreak Alert and Response Network, establishes the human and technical resources to diagnose these outbreaks and thereby check the virus growth. In this review article, we are discussing the outbreaks, precautions along with the appropriate preparedness of individual as well as the government for dealing with these viral diseases.

Comparison of Total Flavonoid, Phenolic Content and Antioxidant Capacity in Leaf and Seed Extracts from White Holy Basil (Ocimum sanctum).

Ocimum sanctum, commonly known as the white holy basil herb belonging to Lamiaceae family is one of the oldest and popular medicinal plant rich in various phytonutrients and antioxidants. In this study, the comparative evaluation of flavonoids, phenolic content, and antioxidant capacity was carried out in methanolic extract prepared from O. sanctum leaves and seeds. The TAC, TPC, and the TFC were measured by ammonium molybdate, Folin-Ciocalteau and aluminum chloride method respectively. Antioxidant activity was also determined by using DPPH and FRAP assay. In response to the above assays, TACs of O. sanctum leaf and seed extracts were 25-248 and 0.011-0.109 μg AAE/10 mg of extract respectively. The TFC assay showed that leaf extract of O. sanctum (14- 225 μg QE/10mg extract) had higher flavonoid content than the seed extract (0.009-0.119 μg QE/10 mg extract) and the TPC assay in the leaf extract (4.49-9.31 μg GAE/mg extract) was higher than those present in seed (4.10-9.05 μg GAE/mg extract). In DPPH assay, % inhibition in O. sanctum leaf extract was determined in the range 18-76% while in seed extract it was 6-29% and in FRAP assay, leaf extract displayed reducing power in range 0.48- 5.50 μg FSE /mg extract while in seed extract it was 0.16-5.46 μg FSE /mg extract. It was observed that O. sanctum leaf extract had high total phenolic and flavonoid content in addition to antioxidant capacity as compared to its seed extract. Abbreviations: TAC: Total Antioxidant Capacity TPC: Total Phenolic Content TFC: Total Flavonoid Content DPPH: 2, 2-diphenyl-1-picryhydrazyl FRAP: Ferric Reducing/Antioxidant Power

Curent advances and stretegies for reducing the side effects of radiation therapies used for various types of cancers.

Cancer originates from the abnormal expression or activation of positive regulators and functional suppression of negative regulators. The World Health Organization (WHO) estimates that 84 million people will die of cancer between 2005 and 2015 without intervention. Research suggests that one-third of cancer deaths can be avoided through prevention. Major cancer treatment modalities are surgery, radiation therapy and chemotherapy. Radiation therapy is an important cancer treatment method and is used for approximately 50% of all cancer patients with varying success. Therapy uses high-energy waves or particles to destroy cancer cells. It can be used basically for three main reasons: to achieve high radiation dose into tumors; minimizing dose into surrounding normal tissues; to avoid complications as far as possible. The recent advances in this treatment method have led to the improvement in cancer death statistics. It can also be combined with surgery or chemotherapy for better results. This review covers general applications, various side effects/agents and factors affecting to get rid of these effects and strategies to improve radiation therapy.