International Journal of Biotechnology Applications

48-51

Authors: HIREMATH K.S., PATIL C.S

This research focused on isolation, purification and characterization of new strain of bacillus, which was able to producing extracellular alkaline thermostable xylanase. Xylanase production by alkalo thermostable Bacillus species were isolated from different regions of Bidar, India and used to screen for the xylanase production by using Xylan plate assay. The paper and sugar industrial waste are used to screen and produce the xylanase, the industrial wastes of paper industry showed the maximum synthesis of xylanase at pH ranging from 9 to 12 and temperature 35 to 70°c after 18-24h. The produced Alkalo thermostable xylanase by Bacillus species showed the most xylanolytic activity at pH >10 and temperature of 70°C.

5-10

Authors: SULTANA S., AKTER S., HOSSAIN M.A.R. , ALAM M.S

The availability of the stinging catfish, Heteropneustes fossilis has been declined in the natural water bodies in Bangladesh in the last two decades. Thus, it is a great concern whether the reduction in abundance has had any impact on genetic variability of this species. Randomly Amplified Polymorphic DNA (RAPD) fingerprint was generated to analyze the genetic variation in five different populations of H. fossilis namely Chalan beel (Natore), Pirgonj beel (Rangpur), Mohongonj haor (Netrakona), Belaboo beel (Narsingdi) and a hatchery located in Mymensingh. Four polymorphic decamer primers, out of 30 tested, were selected for Polymerase Chain Reaction and used in the final RAPD analysis of fifteen individuals from each population. These four primers produced 31 scorable bands of which 26 (83.87%) were polymorphic. The proportion of polymorphic loci and gene diversity estimates were 83.87 and 0.3703, respectively in the Mohongonj population indicating the existence of a relatively higher level of genetic variation in this population. All the populations showed almost similar level of intra-population similarity indices. The UPGMA dendrogram based on Nei’s original measures of genetic distance (D) indicated the segregation of five populations into two distinct groups. The four natural populations made one cluster and the hatchery population produced another cluster. RAPD markers could detect variations within and between populations allowing genetic characterization of different populations of H. fossilis.

20-32

Authors: Kaliwal B.B., Manawadi S.I

Methomyl (S-methyl-1-N [(methyl carbamoyl) oxy] thio acetimidate, is a carbamate broad spectrum insecticide. The Swiss albino mice were administered orally 1, 2, 3, and 4 mg/kg body wt methomyl for 30 days and effective dose 4 mg/kg for 5, 10, and 20 days to know the dose and durational effect on testes and biochemical contents. The mice were sacrificed on day 31st or 24 hours after the terminal exposure. Testes, epididymis, vasa deferentia, seminal vesicles, prostate, coagulatory and Cowper’s glands weight decreased significantly with 2, 3 and 4 mg/kg/day and in 4mg/kg /day for 10 and 20 days of methomyl treatment. Histologic studies of the testes revealed anti-spermatogenic and anti-steroidogenic effects of methomyl reflected by significant decrease in the number and diameter of spermatogonia, primary spematocytes, secondary spermatocytes, spermatids and Leydig cells in the mice treated with 2, 3 and 4 mg/kg/day and in 4 mg/kg /day for 10 and 20 days of methomyl treatment, except with 2 mg/kg/day methomyl treatment number of primary spematocytes, Leydig cells, diameter of spermatogonia, and in 4 mg/kg/day methomyl for 10 days treatment the number of spermatogonia, leydig cells and diameter of primary spermatocytes not changed significantly when compared with controls. Biochemical studies of testis and epididymis showed methomyl at doses of 2, 3 and 4 mg/kg/day and in 4 mg/kg /day for 10 and 20 days of methomyl treatment caused significant decrease in the levels of DNA, RNA, protein, glycogen and sialic acid whereas cholesterol increased significantly. However, treatment with 1, 2 mg / kg/ day and 4 mg / kg /day for 5 days methomyl caused no significant change in epididymal biochemical contents when compared with control. Study on enzyme activities in testis and epididymis revealed that the treatment with 3 and 4 mg/kg/day and 4 mg/kg /day for 20 days of methomyl caused significant decrease in the activities of succinic dehydrogenase (SDH), ions ATPase (Na+-K+ATPase, Ca++ATPase, Mg++ATPase) and acid phosphatase (ACP) were decreased significantly, except with 3 mg/kg/day methomyl treatment where Ca++ ATPase activity in testes and epididymis not changed significantly. However, lactate dehydrogenase (LDH) and alkaline phosphatase (AKP) activities were increased significantly. The testis 3β hydroxysteroid dehydrogenase (3βHSD) decreased significantly with 3 and 4 mg/kg /day and in 4 mg/kg /day for 10 and 20 days of methomyl treatment when compared with controls. However, there was no significant change in testes and accessory reproductive organs, number and diameter of spermatogonia and Leydig cells and biochemical contents of testis and epididymis with 1 mg/kg/day and in 4 mg/kg /day for 5 days of methomyl treatment. These observed effects of methomyl on testes and accessory reproductive organs weight, testis histology, biochemical contents and enzyme activities in testis and epididymis may be due to toxic effect or hormonal imbalance in any of the stage in the hypothalamo-hypophysial-testicular axis.

6-10

Authors: Jaya Barath J., Asma Musfira S., Giridhar R., Arulmurugan R., Shyam Sundar S., Shanmuga Gowtham J

Glycerol finds a lot of industrial applications in the fields of cosmetics pharmaceuticals and in industries. In the present study the production of glycerol is studied using Candida krusei in a batch reactor. The influence of different carbon source on glycerol production is studied using glucose, sucrose and different combinations of glucose and sucrose mixtures. The effect of initial pH was studied by varying the initial pH from 5.0 to 9.0 in order to find the optimum pH to maximize the concentration of cellmass and glycerol. Initial pH 7.0 was found to be an optimum for glycerol fermentation. The maximum glycerol concentration (0.58g/l) was obtained when glucose is used as a carbon source at a concentration of 200g/l.

10-15

Authors: Nikhil Gupta, Soham Trivedi, Hipal Gaudani, Mayank Gupta, Prasad Patil, Girish Gupta, Vamsi Krishna K

Fermentation of citrus juice may offer a relative simple avenue for reducing post-harvest wastage of citrus fruits in low utilization environment and in places where the production of citrus concentrates is low or nonexistent. Orange juice concentrates are readily storable and can be used for production processes even when the fruit is out of season and investigated the possibility of exploiting the fermentative ability of yeasts to produce orange wines.

20-25

Authors: Mishra S.K., Dwivedi S.P., Dwivedi N., Singh R. K., Dubey K.B

Musa accuminata is the member of family Musaceae. The fruits of M. accuminata are harvested in unriped green in order to better withstand transportation and to slow down the natural ripening process because of several weeks travel from the production areas to the end markets. The ripening process is a multi-step reaction involves a number of complex enzymes. The present work is aimed to computational characterization of these ripening enzymes in order to understand the ripening mechanism. In this work identification of the enzymes/proteins involved in ripening process are characterized by their conserved domains (conserved, 3-dimension region). The domains are identified by using the computational tool, CDART (Conserved Domain Architecture Retrieval Tool). By using this applied approach a number of conserved domains were identified, out of these enzymes, the domain pattern of Aspartate aminotransferase (AAT_Like), Glycosyl hydrolase (Glyco_Hydr) and oxoglutarate (20G_Fe) were reported the most conserved followed by Ethylene insensitive (EIN3), GT1, DPBB & Pollen in ripening enzyme complexes. The data, thus, obtained provide new insights in order to understand the role of the proteins playing in different stages of ripening process. These results provide a basis for further studies on the molecular mechanism of ripening process and food technological applications for delayed ripening in fruits, where ripening proteins plays an important role in.

32-40

Authors: Shah R. P., Pawar G. B., Bhiwgade D. A

Adenosine deaminase is a catabolic enzyme, part of the purine salvage metabolic pathway. The principal function of ADA nevertheless appears to be related to the development of immune system and cell differentiation in human. Tuberculosis is an ancient disease of man and is still of the most widespread. Case rates are varied markedly according to age, sex, race and geographic locations. The global incidence of tuberculosis (TB) has sharply increased particularly in areas where HIV and tuberculosis are both prevalent. The Adenosine deaminase activity was found to be elevated in the fluid sample of most of the suspected cases of Tuberculosis and HIV. But it has been observed that though the patients presented typical clinical picture of tuberculosis infection and HIV, the adenosine deaminase activity is much above the reference range. Estimation of the various fractions of protein of the patient with high Adenosine Deaminase activity in the fluid showed increase in albumin, alpha -1 , alpha- 2, beta and gamma globulin while protein electrophoresis of the serum of the same patient showed a decrease in the albumin and increase in the alpha – 1, alpha – 2, beta and gamma globulin. The Adenosine deaminase activity along with the lymphocyte to neutrophil ratio, total leucocyte count and protein electrophoresis can be used as a diagnostic test for the diagnosis of HIV and HIV plus tuberculosis infection.

50-54

Authors: Jayabarath J., Shyam Sundar S., Arulmurugan R., Giridhar R

Industrial waste and sewage pollute more than 2/3 of India’s water resources. Stream pollution is a serious and growing problem in most developing countries where there is little waste water treatment. Increasing contamination of aquatic resources with pollution including heavy metals (like chromium, lead, cadmium, zinc, nickel etc.) endangers aquatic biota and declines water quality. Bioremediation is a process that uses microorganisms or their enzymes to return the environment altered by contaminants to its original condition. Biological methods for the removal of heavy metals from industrial waste may provide an attractive alternative to the physico-chemical process; biosurfactants are one of the compounds that aid in alleviating the heavy metals. Microorganisms while trying to utilize substrates like hydrocarbon as carbon source facilitate the diffusion into cell by producing a variety of substances called biosurfactants. Several microbes such as Bacillus sp., Pseudomonas sp., Acinetiobacter sp. and Arthobacter sp. are reported to produce biosufactants. Compared to synthetic compounds, biosurfactants offer the advantages of little or no environmental impact and the possibility of in situ production. Studies in recent past have demonstrated the successful use of biosurfactants for facilitating the degradation of organic pollutants in soil and water. In the light of above the present study is aimed to carry out the assessment of efficiency of biosurfactants (Rhamnolipid) producing microorganisms (Pseudomonas sp.) isolated from heavy metal contaminated site.

5-13

Authors: Warangkar S.C., Khobragade C.N., Dawane B.S., Bhosale R.B

Purified L-asparaginase in combination with other anticancer drugs like pyrimidine derivatives is administered usually in the body to treat ALL. In the present study, L-asparaginase was purified from Erwinia carotovora up to 247.6 fold and its catalytic properties were studied in the presence of eight different dihydropyrimidine (DHP) derivatives, out of eight derivatives only two viz 4- (2'-hydroxy phenyl) -6- methyl -2- thioxo)-1 -N - benzilydene - 1, 4 - dihydropyrimidine - 5 -carboxylic acid ethyl ester (P1) and 4 -(2'-hydroxy-5'-chlorophenyl)-5-acetyl-6-methyl-2 pyrimidinone (P2) were found to be activators of L-asparaginase. Their catalytic effect was assayed at optimum pH 8.6 and at temperature 35°C in the absence and presence of derivatives P1 and P2 (20-40 μM) at 0.02-0.1 mM concentration of asparagine. It was found that derivatives below the concentration 5 μg/ml have no effect on the activity. Derivative P1 is found to be a strong activator of the asparaginase activity that was reflected by an increase in the Vmax (1.75 fold by P1 and 2.80 fold by P2 respectively) and decrease in the Km (0.91 fold by P1 and 0.81 fold by P2 respectively). The activation of asparaginase is explained by suppressing the cooperativity for the substrate, producing hyperbolic kinetics with Km of 0.080 mM and by 3 fold increase in the Vmax of the enzyme. The activation by derivative P1 and P2 were additive, at optimal or suboptimal concentrations of both activators (up to 30μg/ ml). The DHP derivatives were further analyzed for quantitative SAR study (QSAR) by using PASS, online software to determine their Pa value. Toxicity and drug relevant properties were analyzed by PALLAS software in terms of their molecular weight and log p values. The results showed both the derivatives P1 and P2 are positive modulators of asparaginase activity and may support the development of novel combination therapy for the treatment of Leukemia and solid blood tumors.