Single cell oil production by a novel yeast Trichosporon mycotoxinivorans for complete and ecofriendly valorization of paddy straw

Background: Oleaginous yeasts can be grown on different carbon sources, including lignocellulosic hydrolysate containing a mixture of glucose and xylose. However, not all yeast strains can utilize both the sugars for lipogenesis. Therefore, in this study, efforts were made to isolate dual sugar-utilizing oleaginous yeasts from different sources. Results: A total of eleven isolates were obtained, which were screened for their ability to utilize various carbohydrates for lipogenesis. One promising yeast isolate Trichosporon mycotoxinivorans S2 was selected based on its capability to use a mixture of glucose and xylose and produce 44.86 ± 4.03% lipids, as well as its tolerance to fermentation inhibitors. In order to identify an inexpensive source of sugars, nondetoxified paddy straw hydrolysate (saccharified with cellulase), supplemented with 0.05% yeast extract, 0.18% peptone, and 0.04% MgSO4 was used for growth of the yeast, resulting in a yield of 5.17 g L−1 lipids with conversion productivity of 0.06 g L−1 h−1 . Optimization of the levels of yeast extract, peptone, and MgSO4 for maximizing lipid production using Box­Behnken design led to an increase in lipid yield by 41.59%. FAME analysis of single cell oil revealed oleic acid (30.84%), palmitic acid (18.28%), and stearic acid (17.64%) as the major fatty acids. Conclusion: The fatty acid profile illustrates the potential of T. mycotoxinivorans S2 to produce single cell oil as a feedstock for biodiesel. Therefore, the present study also indicated the potential of selected yeast to develop a zero-waste process for the complete valorization of paddy straw hydrolysate without detoxification

Glycoside hydrolase production by Aspergillus terreus CM20 using mixture design approach for enhanced enzymatic saccharification of alkali pretreated paddy straw.

A successful lignocellulosic ethanol production process needs to address the technological impediments such as cost-competitiveness and sustainability of the process. Effective biomass utilization requires a repertoire of enzymes including various accessory enzymes. Developing an enzyme preparation with defined hydrolytic activities can circumvent the need for supplementing cellulases with accessory enzymes for enhanced hydrolysis. With this objective, mixture design approach was used in the present study to enhance glycoside hydrolase production of a fungal isolate, Aspergillus terreus CM20, by determining the proportion of different lignocellulosic components as enzyme inducers in the culture medium. A mixture of paddy straw and wheat straw (1.42:1.58) resulted in improved cellulolytic activities. The precipitated crude enzyme showed higher CMCase (365.03 18 IU g-1), FPase (161.48 IU g-1), avicelase (15.46 IU g-1), β-glucosidase (920.92 IU g-1) and xylanase (9627.79 IU g-1) activities. The potential of the crude enzyme for saccharification of alkali pretreated paddy straw was also tested. Under optimum conditions, saccharification released 25.0 g L-1 of fermentable sugars. This indicates the superiority of the crude enzyme produced with respect to its hydrolytic enzyme components.

Thermotolerant fermenting yeasts for simultaneous saccharification fermentation of lignocellulosic biomass

Lignocellulosic biomass is the most abundant renewable source of energy that has been widely explored as second-generation biofuel feedstock. Despite more than four decades of research, the process of ethanol production from lignocellulosic (LC) biomass remains economically unfeasible. This is due to the high cost of enzymes, end-product inhibition of enzymes, and the need for cost-intensive inputs associated with a separate hydrolysis and fermentation (SHF) process. Thermotolerant yeast strains that can undergo fermentation at temperatures above 40°C are suitable alternatives for developing the simultaneous saccharification and fermentation (SSF) process to overcome the limitations of SHF. This review describes the various approaches to screen and develop thermotolerant yeasts via genetic and metabolic engineering. The advantages and limitations of SSF at high temperatures are also discussed. A critical insight into the effect of high temperatures on yeast morphology and physiology is also included. This can improve our understanding of the development of thermotolerant yeast amenable to the SSF process to make LC ethanol production commercially viable.

A Comparative Evaluation of the Laryngeal Mask Airway Proseal and Tracheal Intubation for Laparoscopic Cholecystectomy.

The study evaluated 60 adult patients of either sex belonging to ASA grade I & II, who were randomly allocated into 2 groups of 30 each; Group I-Proseal LMA group and Group II -ETT group. They were premedicated with inj. glycopyrrolate 5-10 mcg/kg and inj.butarphanol 2mg intramuscular 45 minutes prior to surgery. Patients were preoxygenated & anaesthesia was induced using oxygen + inj. propofol (2mg/kg) + isoflurane + inj. vecuronium bromide (0.1 mg/kg). PLMA or ETT was inserted as per group. Insertion characteristics i.e. ease of insertion and insertion time were noted. A 16Fr gastric tube was passed into the stomach in every patient and connected to continuous suction. Anesthesia was maintained with nitrous oxide, oxygen, isoflurane and inj. vecuronium bromide. Ventilation was set at 10 ml/kg and 15 breaths/min.The demographic data of both the groups were comparable. The mean age in this study was 42.2 years (22 females and 8 males) in Group I (PLMA) and 40.8 years (23 females and 7 males) in Group II (ETT). The mean BMI of patients in Group I was 22.7 kg/m2 and in Group II was 23.1 kg/m2. Insertion of device was graded as easy in 93% of patients in PLMA group and 96% of patients in ETT group. Mean time taken for successful placement of device in PLMA group was 15.83 s and in ETT group was 17.1 s, which was, however, statistically insignificant (p value = 0.095).The insertion of NGT through drain tube of PLMA was easier than via nose in ETT group. The mean insertion time taken to insert NGT through PLMA was significantly less (10.03 s) than via nose (12.7 s) in intubated patients.The anesthetic and peritoneal insufflation times in both groups were comparable. . The ventilation was adequate to maintain Sp02 of 99% - 100% in both the groups. Based on the study ,it may be said that ,Proseal laryngeal mask airway seemed to be a safe and effective alternative to endotracheal intubation in patients of laparoscopic cholecystectomy.

A new spectrophotometric method for quantification of potassium solubilized by bacterial cultures.

A new spectrophotometric method was developed for the quantification of potassium in the culture broth supernatant of K-solubilizing bacteria. The standard curve of potassium with the new method, which is based on the measurement of cobalt, showed a regression coefficient (R2) of 0.998. The quantification values of potassium obtained with flame photometric method and the newly developed method showed a significant correlation (r) of 0.978. The new method depends on the precipitation of sodium cobaltinitrite with solubilized potassium in liquid medium as potassium sodium cobaltinitrite, which develops bluish green colour by the addition of conc. HCl. The intensity of developed colour can be recorded at 623 nm. This method involves less number of steps, is easy and time saving, and can be used for the reliable estimation of available potassium in culture broth supernatant of K-solubilizing bacteria.

Gastroprotective Activity of Methanol Leaves Extract of Barleria prionitis Linn. on Ethanol and Indomethacin Induced Ulcer in Rats.

Aims: Barleria prionitis L. (Family Acanthaceae) is a medicinal plant found road side in India and whole plant or its various parts like leaves, root, bark, stem and flowers are used traditionally for various treatments like toothache, inflammation, boils, glandular swellings and ulcer. Leaf juice is useful in gastric ulcer. Here, we attempt to prove the use of this plant as gastroprotective agent. Study Design: This study was conducted to evaluate the antiulcer activity of methanol extract obtained from the leaves of Barleria prionitis Linn. Place and Duration of Study: The experiments were conducted at Pharmacology lab of Institute of Pharmaceutical Sciences, Kurukshetra University during the period of July 2012 to December 2012. Material and Methods: Antiulcer activity was performed using the protocols of ulcer induced by ethanol and indomethacin at two different doses (250 and 500mg/kg). Parameters like volume of gastric juice, pH, free acidity, total acidity, aspartate amino transferase (AST) and alanine amino transferase (ALT) were also determined in ethanol induced ulcer model. Results: The reduction in ulcer index in Barleria prionitis treated animals was found to be statistically significant (P=.05), when compared with control groups in both the models. Significant changes were observed in total acidity only at dose 500mg/kg only and changes were significant in AST, ALT levels at both the doses. Other parameters showed non-significant results. Conclusion: The results of the present study show that the methanolic extract of Barleria prionitis L. possess antiulcer activity. This work supports the traditional use of this plant in treating gastric ulcer.

Anti-granuloma activity of Coriandrum sativum in experimental models.

Background: Coriandrum sativum has been used in the traditional systems of medicine for management of arthritis and other infl ammatory disorders. Objectives: In this study, we have evaluated the anti-infl ammatory and anti-granuloma activities of Coriandrum sativum hydroalcoholic extract (CSHE) in experimental models. Materials and Methods: The anti-infl ammatory activity of CSHE was evaluated using carrageenan-induced paw edema model and the anti-granuloma activity of CSHE was evaluated using the subcutaneous cotton pellet implantation-induced granuloma formation and stimulation of peritoneal macrophages with complete Freund’s adjuvant. Serum tumor necrosis factor- (TNF-), IL-6, IL-1  levels, and peritoneal macrophage expression of TNF-R1 were evaluated as markers of global infl ammation. Results: CSHE at the highest dose tested (32 mg/kg) produced a signifi cant reduction (P0.05) in paw edema after carrageenan administration. CSHE treatment also reduced dry granuloma weight in all treated animals. Serum IL-6 and IL-1  levels were signifi cantly (P0.05) lower in the CSHE (32 mg/kg)-treated group as compared to control. Although there was an increase in serum TNF- level in the CSHE-treated group as compared to control, TNF-R1 expression on peritoneal macrophages was found to be reduced. Conclusion: Thus, the result of this study demonstrates the anti-infl ammatory and anti-granuloma activities of CSHE in experimental models, and validates its traditional use for the management of arthritis and other infl ammatory disorders.

Evaluation of disease modifying activity of Coriandrum sativum in experimental models.

Background & objectives: Coriandrum sativum (CS), has been widely used in traditional systems of medicine for treatment of rheumatoid arthritis. However, the mechanism of action for its antiarthritic effects is not clearly known. Therefore, the present study was carried out to evaluate the antiarthritic activity of CS in rats in two experimental models. Methods: The antiarthritic activity of CS seed hydroalcoholic extract (CSHE) was evaluated in adult Wistar rats by using two experimental models, viz. formaldehyde and Complete Freund's adjuvant (CFA) induced arthritis. The expression of pro-inflammatory cytokines (predominantly contributed by macrophages) was also evaluated. TNF-α level was estimated in serum by ELISA method. TNF-R1, IL-1 β and IL-6 expression in the synovium was analysed by immunohistochemistry. Results: CSHE produced a dose dependent inhibition of joint swelling as compared to control animals in both, formaldehyde and CFA induced arthritis. Although there was a dose dependent increase in serum TNF-α levels in the CSHE treated groups as compared to control, the synovial expression of macrophage derived pro-inflammatory cytokines/cytokine receptor was found to be lower in the CSHE treated groups as compared to control. Interpretation & conclusions: Our results demonstrate that the antiarthritic activity of CSHE may be attributed to the modulation of pro-inflammatory cytokines in the synovium. In further studies CSHE could be explored to be developed as a disease modifying agent in the treatment of RA.

Antiarthritic activity of Majoon Suranjan (a polyherbal Unani formulation) in rat.

Background & objectives: Majoon Suranjan (MS) is a polyherbal formulation used in Unani system of medicine for the treatment of rheumatoid arthritis (RA). The present study evaluates the antiarthritic efficacy of this formulation in three different experimental models. Methods: The anti-inflammatory activity of MS (in doses of 450, 900 and 1800 mg/kg body wt) was evaluated using the turpentine oil induced paw oedema model and the antiarthritic efficacy was evaluated using the formaldehyde and complete Freund's adjuvant (CFA) induced arthritis models. Aspirin (100 mg/kg body wt) was used as the standard drug in all the models. In order to assess the safety of the test drug, oral acute and 28 day toxicity studies were also carried out. Results: MS produced a dose dependent protective effect in all the experimental models. Its antiarthritic efficacy was comparable to aspirin in formaldehyde induced arthritis and was superior to aspirin in turpentine oil induced paw oedema and CFA induced arthritis. MS also inhibited the delayed increase in joint diameter as seen in control and aspirin treated animals in CFA induced arthritis. Oral LD50 of MS was found to be >5000 mg/kg in rats. Chronic administration did not produce any significant physiological changes in the tested animals. Interpretation & conclusions: Results of the present study suggest that the antiarthritic activity of MS was due to the interplay between its anti-inflammatory and disease modifying activities, thus supporting its use in traditional medicine for the treatment of RA.

Evaluation of anti-inflammatory activity of plant lipids containing alpha-linolenic acid.

Two groups of fatty acids are essential to the body, the omega6 (n6) series derived from linoleic acid (18:2, n-6) and the omega3 (n3) series derived from alpha-linolenic acid (18:3, n-3). Fatty acids provide energy, are an integral part of the cell membranes and are precursors of prostaglandins, thromboxanes and leukotrienes collectively known as eicosanoids. Eicosanoids participate in development and synthesis of immunological and inflammatory responses. The fixed oils (1, 2, 3 ml/kg) containing alpha-linolenic acid, obtained from the seeds of Linseed (Linum usitatissimum), Soyabean (Glycine max) and Holy basil (Ocimum sanctum) were screened for their antiinflammatory activity using carrageenan, leukotriene and arachidonic acid induced paw edema models in rats and the antiinflammatory effects were compared with the standard drug indomethacin. Significant inhibition of paw edema was produced by all the oils in the highest dose (3 ml/kg) in all the models. While O. sanctum oil produced the maximum percentage inhibition in leukotriene induced paw edema, L. usitatissimum oil produced maximum percentage inhibition in carrageenan and arachidonic acid induced paw edema models. The results show that oils with higher alpha-linolenic acid content (L. usitatissimum and O. sanctum) produced a greater inhibition of paw edema suggesting that modulation of the course of inflammatory disorders may be achieved by altering the eicosanoid precursor (i.e. poly unsaturated fatty acids: PUFA) availability through dietary manipulation.

Attenuation of the effect of lindane on immune responses and oxidative stress by Ocimum sanctum seed oil (OSSO) in rats.

Present study was done to evaluate the effect of Ocimum sanctum seed oil (OSSO) on the immunotoxicity and oxidative activity of lindane in rats. Rats were divided into four groups (n = 8) and were treated with lindane (10 mg/kg, po) and/or OSSO (1 mg/kg, po) during the study period. Humoral immunity was assessed by measuring haemagglutination titre to sheep red blood cells (SRBC) and delayed type hypersensitivity (DTH) was assessed by measuring foot pad thickness. Lindane showed significant decrease in anti-SRBC antibody titre and also decreased percentage change in foot pad thickness in DTH response as compared to control group. OSSO per se produced significant increase in anti-SRBC antibody titre, but did not produce significant change in the foot pad thickness as compared to control group. However, it significantly antagonized the effect of lindane on the anti-SRBC antibody titre and foot pad thickness parameters. Lindane produced oxidative stress as indicated by increase in the levels of MDA and decrease in GSH levels. Treatment with OSSO per se showed antioxidant activity and also reversed the oxidative stress produced by lindane. The results suggest that OSSO can attenuate the immunotoxicity and oxidative stress produced by lindane.

Biological activities of Ocimum sanctum L. fixed oil--an overview.

Seeds of Ocimum sanctum L. (Labiatae; popularly known as 'Tulsi' in Hindi and 'Holy Basil' in English) contain a pale yellow colored fixed oil. The oil possesses antiinflammatory activity due to dual inhibition of arachidonate metabolism supplemented by antihistaminic activity. The antiinflammatory activity is not dependent on the pituitary adrenal axis. The oil possesses antipyretic activity due to prostaglandin inhibition and peripherally acting analgesic activity. The oil has been found to be effective against formaldehyde or adjuvant induced arthritis and turpentine oil induced joint edema in animals. Lipoxygenase inhibitory, histamine antagonistic and antisecretory activities of the oil contribute towards antiulcer activity. The oil can inhibit enhancement of vascular capillary permeability and leucocyte migration following inflammatory stimulus. The LD50 of the oil is 42.5 ml/kg and long-term use of oil at 3 ml/kg dose does not produce any untoward effects in rats. The oil contains a-linolenic acid, an omega-3 fatty acid, which on metabolism produces eicosapentaenoic acid and the same appears to be responsible for the biological activity. The oil has hypotensive, anticoagulant and immunomodulatory activities. Antioxidant property of the oil renders metabolic inhibition, chemoprevention and hypolipidaemic activity. Presence of linolenic acid in the oil imparts antibacterial activity against Staphylococcus aureus. The oil alone or in combination with cloxacillin, a beta-lactamase resistant penicillin, has been found to be beneficial in bovine mastitis, an inflammatory disorder resulting from staphylococcal infection. Existence of anti-inflammatory, analgesic and antibacterial activities in single entity i.e. fixed oil appears to be unique.

Antidiabetic, antihypercholesterolaemic and antioxidant effect of Ocimum sanctum (Linn) seed oil.

Antihyperlipidaemic and antioxidant effect of Ocimum sanctum Linn. seed oil (OSSO) was investigated in rabbits. Administration of OSSO (0.8 g/kg body weight/day) for four weeks, in cholesterol (100 mg/kg body weight/day) fed rabbits significantly decreased serum cholesterol, triacylglycerol and LDL-+VLDL-cholesterol as compared to untreated cholesterol fed group. There was significant fall in atherogenic index in OSSO treated group. In addition, treatment with OSSO decreased lipid peroxidation and increased reduced glutathione content in blood. Antidiabetic effect of O. sanctum seed oil was evaluated in alloxan diabetic rabbits. Two weeks treatment of diabetic rabbits with OSSO (0.8 gm/kg/day) showed no significant hypoglycaemic effect. Results of the present study show that OSSO has hypocholesterolaemic and antioxidant effects but it does not have antidiabetic effect.

Antibacterial activity of Ocimum sanctum L. fixed oil.

Ocimum sanctum fixed oil showed good antibacterial activity against Staphylococcus aureus, Bacillus pumilus and Pseudomonas aeruginosa, where S. aureus was the most sensitive organism. Sesame and soyabean oils also showed moderate activity against S. aureus. Higher content of linolenic acid in O. sanctum fixed oil could contribute towards its antibacterial activity. The antibacterial activity combined with anti-inflammatory and analgesic activities of the oil, could make it useful in inflammatory disorder resulting from staphylococcal infection.