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.

To Evaluate Efficacy of Sonographic Cervical Length to Predict Preterm Labour.

Background: Preterm birth is a major challenges faced by obstetricians worldwide during their practice. Preterm birth is one of a leading direct cause of neonatal death (27%) and more than one million preterm newborns die annually due to various complications. This study done to look for diagnostic efficacy of sonographic cervical length to predict preterm labour. Methods: A prospective study comprised of a total of 132 pregnant women with a singleton foetus with symptoms of preterm labour. Sonographic trans-cervical length measurement taken in all patients. Results: Out of which 17 patients were lost during follow up, 3 patients develop premature rupture of membrane. Hence, a study was conducted over 112 patients. Among these 62 patients went in preterm labour and 50 patients delivered at term. For predicting preterm delivery, sensitivity, specificity, positive and negative predictive value of cervical length was 82.2%, 74%, 79.6% and 77%. Conclusion: Sonographic cervical length is a good predictor. Patients with reduced cervical length are more likely to deliver preterm.

Massive Fetomaternal Hemorrhage.

Massive fetomaternal hemorrhage (FMH) is a rare entity and the diagnosis needs to be determined by performing a Kleihauer- Betke test (KBT) of the maternal blood. We present a case of massive FMH presenting as severe neonatal anemia. The varied presentations of FMH and its management are discussed.

Psychogenic excoriation responding to fluoxetine: a case report.

Compulsive skin picking, 'acne excoriee', neurotic (psychogenic) excoriation, dermatotillomania, occurring in 2% dermatology patients mostly in women, is a result of excessive scratching, picking, gouging or squeezing of the skin using teeth, tweezers, nail files, pins and knives, etc. The lesions are usually found on face and also on upper limbs and upper back, areas patients can easily reach. They may occur in absence or in response to skin pathology or sensation of itching. A young female patient attended OPD with the complaints of multiple excoriated lesions over the face, arms and forearms. The diagnosis was psychogenic excoriation which is an uncommon psychodermatological condition. She was treated with fluoxetine and behaviour therapy. The patient recovered fully with above treatment at the end of 3 months. Psychogenic excoriation is an uncommon psychodermatological condition which responds well to selective serotonin reuptake inhibitors and behaviour therapy (habit reversal training).