Search

Cellulose, a carbohydrate mainly found in plant cell walls, is most abundant biopolymer on the Earth. Biodegradation of cellulose is carried out by a group of enzymes knows as cellulases. These enzymes play important roles in food processing and used in food industry. In the present era, cold-active enzymes are preferred instead of meso- and thermos-philic counterparts due to less energy requirement for their optimal activity and easy inactivation. The present study includes evaluation of cold-active cellulase from Pseudoalteromonas haloplanktis for its industrial applications in comparison to mesophilic and thermophilic cellulases, through molecular docking method. The binding energy of cold-active cellulase with the substrate cellulose was -126.60 KCals/mole. However, the energy for thermo- and mesophilic cellulase found to be -93.29 and -75.54 KCals/mole, respectively. The results concluded that cold-active cellulase has more efficacy compared to its counterparts and may be used in food processing industry at commercial level.

Cellulose is the most bountiful organic compound in the world and a major renewable agriculture waste with exceptionally high potential for biological alteration into valuable bioproducts. Cellulose also serves as a good source of low-cost energy production in the form of biofuels. Cellulase is the broad name for the group of three enzymes, namely endo-1-4-β-glucanase (EC 3.2.1.4), cellobiohydrolase (EC 3.2.1.91), and β-glucosidase, which catalyze the hydrolysis of cellulose and related oligosaccharides. Microorganisms and plants commonly synthesize such molecules. Cellulases fall into the main group of industrial enzymes that have extensive applications in many areas, such as the production of ethanol and organic acids, cotton processing, paper recycling, as animal feed additives, for brightness of fiber surfaces in paper industries, to enhance pulp drainage in paper mills, etc. However, the largest obstacle for the commercialization of these enzymes is their high manufacturing price, which can be minimized by using other carbon sources, for example, agricultural solid waste residues during optimization of the production medium. This chapter summarizes the applications of cellulase enzymes along with their sources but mainly focuses on the various bioprocess parameters used for production optimization, including the latest technologies/techniques applied at an industrial scale.

The present study was accomplished to examine and compare the effect of specific antioxidant-rich oils on hyperglycemia, dyslipidemia, renal function markers and oxidative renal damage in diabetic rats for four weeks. Papaya (P), olive (O), fenugreek (Fe), bitter gourd (B) and fish (Fi) oils were used for this purpose. Streptozotocin (STZ) was injected intraperitoneally in a single dose to induce diabetes. All oils were given orally at a dose of 3g/kg for four weeks in respective group after induction of diabetes. After treatment with oils, blood was collected, and their kidneys were stored. The level of fasting blood glucose (FBG), glycated hemoglobin (HbA1c), total cholesterol (TC), triglycerides (TG), low-density lipoprotein-cholesterol (LDL-C) and very low-density lipoprotein-cholesterol (VLDL-C) increased while amylase and high-density lipoprotein cholesterol (HDL-C) level decreased in the diabetic rats. These changes were augmented by fenugreek, bitter gourd and olive oils treatment. Diabetic rats showed elevated renal function markers in serum, including, serum creatinine (Scr), blood urea nitrogen (BUN) and alkaline phosphatase (ALP), which were restrained significantly by fenugreek and bitter gourd oil treatment. Moreover, fenugreek and bitter gourd oils treatment significantly modulated the level of thiobarbituric reactive substances (TBARS), malonaldehyde (MDA) and catalase (CAT) in the kidney of diabetic rats. The histopathological examination also showed the protective effect of these oils. The study suggests that vegetable oils are effective in reducing hyperglycemia, dyslipidemia and renal damage related to the side effects of diabetes. Thus they may have therapeutic value for preventing diabetes side effects and may be included in oil diet treatment synergically. Thus, our data suggest that oils as potent antidiabetic agent and beneficial in the control of diabetes-related abnormalities such as hyperglycemia, dyslipidemia and renal damage of STZ induced rat model of type 2 diabetes. Our study also supports the suggestion that synergistic possibilities exist concerning the use of these oils in the treatment of diabetes mellitus.

Enzymes are the chemical compounds which act as a biological catalyst and alter the rate of biochemical reactions. There are about 3000 enzymes in our bodies which are involved in more than 7000 metabolic reactions. Considering the physiological and metabolic significance of the enzymes, food enzymes constitute an important part of diet. There are some enzyme-rich foods that are believed to have a different function in the human body than that of body’s own enzymes. But, processing and cooking of foods completely degrade the enzyme contents and leave the body deprived of these important enzymes which play a significant role in the well-being and healthcare. Some prominent examples of high enzyme foods are papaya, pineapple, banana, figs, and bee pollen. The sprouts are metabolically very active, and the content of enzymes is more as compared to seeds. The dietary enzymes commonly present in the enzyme-rich foods are proteinases, amylases, maltases, lipases, papain, bromelain, etc. Non-processed and uncooked foods are rich in enzymes, and their intake might decrease the body’s burden to produce more and more of its own enzymes. As a result of some metabolic disorders and diseases, the production of some specific enzymes is hampered in the body. In such cases, foods which are high in enzyme contents are the best choice for the extraction, purification, and commercialization of the enzymes. These enzymes find numerous applications in pharmaceutical industries, and there is a need for exploiting these foods for industrial applications.

Microbacterium foliorum GA2, an alkali-tolerant bacterium, was randomly mutated using UV radiation and sodium azide to obtain a mutant with a higher cold-active extracellular amylolytic activity. A mutant, designated as MFSD20, was selected owing to its higher amylase activity at 20°C. Under optimized conditions, amylase production was achieved best with raw banana peels (5000 units) in solid-state fermentation (SSF). The enzyme was purified by salt precipitation and chromatographic methods and afterwards characterized biochemically. The purified enzyme showed maximal activity at temperatures between 15-25°C and at pH 8.0. Interestingly, this mutant biocatalyst (MFSD20) displays higher catalytic activity under conditions of low temperature (4°C) and high pH (10.0), in the presence of SDS (0.1 and 1%), and exhibited 85% and 50% requirement of divalent metallic ions Ca2+ and Mg2+, respectively. This mutant enzyme extract in combination with "Wheel detergent" was highly effective in the removal of tomato sauce and chocolate stains from white cotton fabric was demonstrated by ~50% additional reflectance compared with detergent alone, in a wash performance analysis at 20 ± 2°C. The features shown by mutant M. foliorum GA2 make it a promising candidate for industrial applications involving starch degradation at low temperatures.

In the present study, four cold-adapted bacterial isolates were screened for multiple-enzyme production at low temperature (15 °C). The most potent isolate, Bacillus cereus GA6 (HQ832575), was subjected to mutation by UV radiation to obtain a mutant strain with elevated enzyme production. The mutant strain, designated as CUVGA6, with higher chitinase activity at low temperature was selected for enzyme production optimization using factorial design and response-surface methodology (RSM). Two statistically significant parameters (colloidal chitin and KH2PO4) for response were selected (p value = 0.008 and 0.004, respectively) along with pH and temperature and utilized to optimize the process. Central composite design of RSM was used to optimize the levels of key ingredients for the best yield of chitinase. Maximum chitinase production was predicted to be 428.57 U/ml for a 4.4-fold increase in medium containing 2 % colloidal chitin, 6.0 g/L K2HPO4 and pH 9.0 at 25 °C when incubated for 7 days in submerged fermentation. ANOVA of CCD suggested that the quadratic interaction effect of K2HPO4 with chitin, temperature and pH has high impact on the production of chitinase (p value = 0.007, 0.002, 0.035, respectively), although its linear effect was not significant as observed. The closeness of optimized values (R 2 = 82.28 %) to experimental values (R 2 = 80.13 %) proved the validity of statistical model. Thus, multi-enzyme producing cold-adapted mutant B. cereus GA6 (CUVGA6) could be exploited for the production of chitinase which is of industrial significance.

Cold-active enzymes along with their producing microbes are of commercial value and find multiple applications in various industrial and biotechnological sectors. In this study, production optimization of cold active α-amylase from cold-adapted Microbacterium foliorum GA2 was carried out. The aim of the present work was to use economical agro-substrate for increasing cold-active α-amylase production and to optimize the fermentation parameters in SSF (Solid state fermentation) using two statistical experimental designs. Plackett–Burman design and response surface methodology was used to determine key ingredients for the best media composition and optimal concentration of these components, respectively. The screening result of Plackett–Burman design showed bagasse, lactose and pH had significant effects ( p -value≤0.05) on cold-active α-amylase production. Maximum α-amylase production (6610 units) was observed through response surface methodology in medium having 40% bagasse, 0.003 M lactose, and pH 8.0 at 20 °C when incubated for 5 days in static conditions. The closeness of optimized values ( R 2 =92.26%) to experimental values ( R 2 =96.28%) proved the validity of the statistical model. Under these experimental designs, the α-amylase yield increased three-fold in comparison to control and was much efficient and economical than “one-variable-at-a-time” methodology. Thus, cold-adapted M. foliorum GA2 could be exploited for industrial production of α-amylase using bagasse at relatively low temperatures.

A novel laccase enzyme producing bacterium, Pseudomonas putida MTCC 7525, was isolated and subjected to optimization of laccase production. Maximum production (94.10 U/ml) was achieved at 30 °C and pH 8 (108 h incubation) with 10% skim milk and 1 mM sodium nitrate as additional nitrogen source. The laccase was purified by salt precipitation followed by ion exchange chromatography which showed 6.44 fold purification. The purified enzyme had optimal activity at pH 8.0 and 40 °C, and showed stability in DMSO retaining more than 85% of original activity. In presence of manganese and cadmium, enzyme retained >95% and 92% activity, respectively. The molecular weight of laccases was 39.5 kDa and activity was inhibited by pCMB (93%). The synthetic dyes (0.02%) and industrial effluents (10%) were decolorized to 74–93 and 58–68%, respectively when treated with culture of P. putida . However, the culture supernatant of P. putida showed about 36–94 and 16–86% decolorization of synthetic dyes and effluents, respectively within 24 h of incubation.

Chitin is one of the most abundant biopolymers widely distributed in the marine and terrestrial environments. Chitinase enzyme has received increased attention due to its wide range of biotechnological applications, especially in agriculture for biocontrol of phytopathogenic fungi and harmful insects. In the present study, 58 bacterial isolates were screened for chitinolytic activity and on the basis of chitin hydrolysis zone 6 isolates were selected for chitinase production in broth media. Based on enzyme production, two most potent isolates identified as Aeromonas hydrophila HS4 and Aeromonas punctata HS6 were selected for further study. The effects of media composition and various fermentation conditions for optimization of chitinase production were studied. The maximum chitinase production was obtained at 37 °C and pH 8.0 after 24–48 h of incubation by HS4; and at 37 °C and pH 7 after 48 h incubation by HS6. Among the substrates colloidal chitin was the best for both the strains. Regarding carbon sources, starch (1%) was the best for both strains; while malt and yeast extract (1%) was found as the best nitrogen source for HS4 and HS6, respectively. Out of metal ions Mn 2+ and Cu 2+ enhanced enzyme production in the case of HS6. However, Co 2+ was the most appropriate for HS4.

The recent studies carried out on the binding of small molecule to deoxyribonucleic acids suggested that the intercalation of a tricyclic heteroaromatic molecule, thionine, with natural DNAs provided thermal stabilization to the DNA complex. In the present study, we reported theoretical analysis of thionine binding with natural DNAs of varying base composition by using an amended Zimm and Bragg theory, to explain the melting behaviour and heat capacity of DNAs with and without thionine binding. We used experimental models of Paul et al. for implementing this study (Paul et al., 2010). The sharpness of transition has been examined in terms of half width and sensitivity parameter (ΔH/σ). The results of theoretical analysis concluded that the various parameters such as heat capacity curve, transition profile, half widths and sharpness of the transition are in good agreement with the experimental measurements for binding of thionine determined through DSC. The theoretical analysis proposed in this study, therefore, may be useful to understand interaction of small molecules with deoxyribonucleic acids. This approach may also be applied to design DNA binding therapeutic molecules and in the process of drug formulation and development.

Due to high level of blood sugar, some glucose molecules are attached to the normal hemoglobin which is known as glycated hemoglobin (HbA1c). HbA1c indicates the average blood glucose levels over the past 6 to 8 weeks. Therefore, it has been recommended as a screening tool for the detection and monitoring of diabetes mellitus. HbA1c at high levels are associated with various metabolic disorder and other disease. In the present review, we summarized risk factors associated with increased level of HbA1c and its clinical significance.

Microbial cold-active α-amylases offer various economical and ecological benefits through energy savings by overcoming the heating requirements and also provide large biotechnological potentials. The objective of present study was to isolate new cold-adapted bacterial strains for production of cold-active α-amylases and their production optimization. Out of 30 cold-active α-amylase producing bacteria, isolated from soil of Gangotri glacier, Western Himalaya, India, two potential isolates, designated as GA2 and GA6, were selected for enzyme production. The α-amylase production was found maximum at 20 °C and pH 9 after 120 h incubation for GA2; and 20 °C and pH 10 after 96 h incubation for GA6. Among the carbon sources, lactose and glycerol was most suitable for GA2 and GA6, respectively. However, yeast extract and ammonium acetate was found best as nitrogen source by GA2 and GA6, respectively. Out of two potential isolates, maximum enzyme production (5870 units) was achieved with GA2 followed by GA6 (4746 units). GA2 was resistant to penicillin (10 μg) among tested antibiotics and as per plasmid curing results, amylase production was a plasmid mediated characteristic. The phylogenetic analysis revealed that GA2 and GA6 have highest homology with Microbacterium foliorum (99%) and Bacillus cereus (98%), respectively. This was the first report on cold-active α-amylase production by M. foliorum strain GA2 and B. cereus strain GA6, also their 16S rRNA sequences assigned an accession number HQ832574 and HQ832575 , respectively from NCBI.

Biodetergents are preferred over the conventional synthetic detergents in view of their better cleaning properties, low energy input and the alleviation of pollution. The biodetergents derived from mesophilic/thermophilic organisms and also peroxide-based synthetic detergents require high temperature for their optimum activity. Thus, cold-active enzymes are very useful as they work at lower temperatures and do not require the input of energy. The purpose of the present study was the production optimization and purification of cold-active alkaline protease from a novel psychro-tolerantStenotrophomonas maltophilia MTCC 7528 and its application as a detergents additive for cold washing. Psychro-tolerant proteolytic bacterium S. maltophilia MTCC 7528 was isolated from soil of Gangotri glacier, Western Himalaya, India that produced maximum protease (56.2 U/ml) at 20°C and pH 9.0 after 120 h incubation in shaking condition (120 rev/min). The purified enzyme has molecular weight of 75 kDa with maximum activity and stability at pH 10 and 20°C temperature. It showed excellent compatibility with commercial detergents with improved cleansing power at low temperature. The enzyme completely removed blood and grass stains and increases the reflectance by 26 and 23%, respectively. Enzyme-based detergents find a wide range of applications in laundry and textile industries. Cold-active alkaline protease from psychro-tolerant S. maltophilia may be a potential component to be used as a detergent additive for cold washing that will be beneficial to save energy as they work at lower temperatures. Key words: Alkaline protease, biodetergent, cold-active enzymes, wash performance.

Amylases have most widely been reported to occur in microorganisms, although they are also found in plants and animals. Microbial cold-active α-amylases confer high activities at low temperature that favours production of comparatively insubstantial compounds. Cold-active α-amylases have an advantage under extreme low temperature conditions due to their freeze tolerance mechanism, intrinsic greater membrane fluidity and production of cold-acclimation proteins. Due to considerable progress towards energy savings process in industries, the low temperature stability has been regarded as the most important characteristics of cold-active amylases. Cold active enzymes attract more and more attention nowadays. Comparatively little information is available on cold-active microbial α-amylases and its catalytic properties. Now the situation is changing which recently enchanted the scientific community to focus in various fields, such as analytical, medicinal and clinical chemistry, as well as their extensive industrial applications such as starch industry, food processing, additive in detergents, waste-water treatment, biopulping, textile industry, environmental bioremediation in cold climates and other molecular biology applications. Key words: Psychrophiles, psychrotrophs, extremozymes, cold-active amylase, starch degrading enzymes.

A novel psychro-tolerant bacterium, Curtobacterium luteum, secreting an extracellular protease was isolated from the soil of Gangotri glacier, Western Himalaya. Maximum enzyme production was achieved when the strain was grown in a pH-neutral medium containing skim milk at 15 degrees C over 120 h. The metal ions such as Zn(2+) and Cr(2+) enhanced enzyme production. The specific activity of purified enzyme was 8,090 units/mg after 34.1-fold purification. The 115 kDa enzyme was a metalloprotease (activity inhibited by EDTA and EGTA) and showed maximum activity at 20 degrees C and pH 7. The enzyme was active over a broad pH range and retained 84% of its original activity between pH 6 and 8. There was no loss in enzyme activity when exposed for 3 h at 4-20 degrees C. However, the enzyme lost 65% of activity at 30 degrees C, and was almost inactivated at 50 degrees C, but was resistant to repeated freezing and thawing. The enzyme activity was stimulated by manganese ions; however, it was inactivated by copper ions.