Fermentation of Germinated Native Black Rice Milk Mixture by Probiotic Lactic Acid Bacteria

This research aimed to demonstrate probiotic germinated native black rice juice fermentation by lactic acid bacteria (Lactobacillus casei TISTR 390). Germinated native black rice juice was inoculated with a 24-h old lactic culture and incubated at 30 °C for 72 hours. Changes in pH, acidity, total soluble solid, and viable cell counts during fermentation under controlled conditions at 0-h, 24-h, 48-h, and 72-h fermentations were evaluated. The study found out that the change in pH and total soluble solid of probiotic germinated black rice juice significantly (p ≤ 0.05) decreased at 72-h fermentation (5.67±0.12 to 2.86±0.04 and 7.00±0.00 to 6.40±0.00 ºbrix at 0-h and 72-h fermentations, respectively). On the other hand, the amount of titratable acidity expressed as lactic acid and the viable cell count significantly (p≤0.05) increased at 72-h fermentation (0.11±0.06 to 0.43±0.06 (% lactic acid) and 3.60 x 106 to 2.75 x 108 CFU/ml at 0-h and 72-h fermentations, respectively). Interestingly, the amount of γ-Amino Butyric Acid (GABA) had a significant difference (p≤0.05) twice as high as that of the control group (0.25±0.01 and 0.13±0.01 mg/100g, respectively). In addition, the free radical scavenging capacity assayed by DPPH method also showed that the IC50 values were significantly (p≤0.05) higher than the control (147.71±0.96 and 202.55±1.24 mg/ml, respectively). After 4 weeks of cold storage at 4 °C, the viable cell counts of lactic acid bacteria reduced to 1.37 x 106 CFU/ml. In conclusion, fermented germinated native black rice juice could be served as a healthy beverage for vegans and people who are allergic to cow milk products.

Influence of Environment-Friendly Organic Wastes on the Properties of Sandy Soil under Growing Zea mays L. in Arid Regions

Environment-friendly organic wastes of Brewers' spent grain, a byproduct of the brewing process, have recently used as soil amendment to improve soil fertility and plant production. In this work, treatments of 1% (T1) and 2% (T2) of spent grains, 1% (C1) and 2% (C2) of compost and mix of both sources (C1T1) were used and compared to the control for growing Zea mays L. on sandy soil under arid Mediterranean climate. Soils were previously incubated at 65% saturation capacity for a month. The most relevant soil physical and chemical parameters were analysed. Water holding capacity and soil organic matter (OM) increased significantly along the treatments with the highest values in T2. Soil pH decreased along the treatments and the lowest pH was in C1T1. Bicarbonate decreased by 69% in C1T1 comparing to control. Total nitrogen (TN) and available P varied significantly among all treatments and T2, C1T1 and C2 treatments increased 25, 17 and 11 folds in TN and 1.2, 0.6 and 0.3 folds in P, respectively related to control. Available K showed the highest values in C1T1. Soil micronutrients increased significantly along all treatments with the highest values in T2. After corn germination, significant variation was observed in the velocity of germination coefficients (VGC) among all treatments in the order of C1T1>T2>T1>C2>C1>control. The highest records of final germination and germination index were in C1T1 and T2. The spent grains may compensate deficiencies of macro and micronutrients in newly reclaimed sandy soils without adverse effects to sustain crop production with a rider that excessive or continuous use need to be circumvented.

Effects of Gamma Radiation on Tomato Leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

In present study, it was aimed to evaluate the gamma radiation impacts on tomato leaf miner at different biological stages. The laboratory colony of tomato leaf miner was used to set up the experiments. Different biological stages of the insects (eggs, 4th instars and pupae) were irradiated using Cobalt-60 at doses of 0 (control), 100 Gray (Gy), 200 Gy, 300 Gy and 400 Gy in Cos-44HH-N source, at dose rate of 480 Gy/h. After irradiation, the eggs were incubated until hatching; the mature larvae were reared to complete their developments. Adult emergences from irradiated pupae were also evaluated. The results showed that there were no egg hatching at all tested irradiation doses. Although, the pupal percentages of irradiated mature larvae were 54%, 15% and 8% at doses of 100 Gy, 200 Gy and 300 Gy respectively, there were no adult emergences from irradiated mature larvae. On the other hand, the adult emergences were observed from irradiated pupae, decreased as radiation doses increased along with malformed adult appearance. Male and female individuals were out crossed with laboratory reared adults. Fecundity was correlated with radiation doses.

Degradation of Endosulfan in Different Soils by Indigenous and Adapted Microorganisms

The environmental fate of organic contaminants in soils is influenced significantly by the pH, texture of soil, water content and also presence of organic matter. In this study, biodegradation of endosulfan isomers was studied in two different soils (Soil A and Soil B) that have contrasting properties in terms of their texture, pH, organic content, etc. Two Nocardia sp., which were isolated from soil, were used for degradation of endosulfan. Soils were contaminated with commercial endosulfan. Six sets were maintained from two different soils, contaminated with different endosulfan concentrations for degradation experiments. Inoculated and uninoculated mineral media with Nocardia isolates were added to the soils and mixed. Soils were incubated at a certain temperature (30 °C) during ten weeks. Residue endosulfan and its metabolites’ concentrations were determined weekly during the incubation period. The changes of the soil microorganisms were investigated weekly.

Evaluation of Hazelnut Hulls as an Alternative Forage Resource for Ruminant Animals

The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; MEGP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.

Management Control Systems in Post-Incubation: An Investigation of Closed Down High-Technology Start-Ups

Insufficient informal communication systems can lead to the first crisis (‘Crisis of Leadership’) for start-ups. Management Control Systems (MCS) are one way for high-technology start-ups to successfully overcome these problems. So far the literature has investigated the incubation of a start-up, but focused less on the post-incubation stage. This paper focuses on the use of MCS in post-incubation and, if failed start-ups agree, on how MCS are used. We conducted 14 semi-structured interviews for this purpose, to obtain our results. The overall conclusion is that the majority of the companies were closed down due to a combination of strategic, operative and financial reasons.

Evaluation of the Beach Erosion Process in Varadero, Matanzas, Cuba: Effects of Different Hurricane Trajectories

The island of Cuba, the largest of the Greater Antilles, is located in the tropical North Atlantic. It is annually affected by numerous weather events, which have caused severe damage to our coastal areas. In the same way that many other coastlines around the world, the beautiful beaches of the Hicacos Peninsula also suffer from erosion. This leads to a structural regression of the coastline. If measures are not taken, the hotels will be exposed to the advance of the sea, and it will be a serious problem for the economy. With the aim of studying the intensity of this type of activity, specialists of group of coastal and marine engineering from CIH, in the framework of the research conducted within the project MEGACOSTAS 2, provide their research to simulate extreme events and assess their impact in coastal areas, mainly regarding the definition of flood volumes and morphodynamic changes in sandy beaches. The main objective of this work is the evaluation of the process of Varadero beach erosion (the coastal sector has an important impact in the country's economy) on the Hicacos Peninsula for different paths of hurricanes. The mathematical model XBeach, which was integrated into the Coastal engineering system introduced by the project of MEGACOSTA 2 to determine the area and the more critical profiles for the path of hurricanes under study, was applied. The results of this project have shown that Center area is the greatest dynamic area in the simulation of the three paths of hurricanes under study, showing high erosion volumes and the greatest average length of regression of the coastline, from 15- 22 m.

An Antibacterial Dental Restorative Containing 3,4-Dichlorocrotonolactone: Synthesis, Formulation and Evaluation

The objective of this study was to synthesize and characterize 5-acryloyloxy-3,4-dichlorocrotonolactone (a furanone derivative), use this derivative to modify a dental restorative, and study the effect of the derivative on the antibacterial activity and compressive strength of the formed restorative. In this study, a furanone derivative was synthesized, characterized, and used to formulate a dental restorative. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed restorative. The fabricated restorative specimens were photocured and conditioned in distilled water at 37oC for 24 h, followed by direct testing for CS or/and incubating with S. mutans for 48 h for antibacterial testing. The results show that the modified dental restorative showed a significant antibacterial activity without substantially decreasing the mechanical strengths. With addition of the antibacterial derivative up to 30%, the restorative kept its original CS nearly unchanged but showed a significant antibacterial activity with 68% reduction in the S. mutans viability. Furthermore, the antibacterial function of the modified restorative was not affected by human saliva. The aging study also indicates that the modified restorative may have a long-lasting antibacterial function. It is concluded that this experimental antibacterial restorative may potentially be developed into a clinically attractive dental filling restorative due to its high mechanical strength and antibacterial function.

In vitro Environmental Factors Controlling Root Morphological Traits of Pineapple (Ananas comosus L. Merr)

Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Microbiological Profile of UTI along with Their Antibiotic Sensitivity Pattern with Special Reference to Nitrofurantoin

Urinary Tract Infections are considered as one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to wide spread usage of uncontrolled antibiotics. In this study, a total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G. M. C Patiala (January 2013 to December 2013). Urine samples were inoculated on MacConkey’s and blood agar plates and incubated at 370C for 24 hrs. The organisms were identified by colony characters, Gram’s staining, and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby Bauer DISK diffusion method on Muller Hinton agar plates. Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Culture was positive in 25% of the samples. Escherichia coli was the most common isolate 60.3% followed by Klebsiella pneumoniae 13.5%, Proteus spp. 9% and Staphylococcus aureus 7.6%. Most of the urinary isolates were sensitive to, carbepenems, Aztreonam, Amikacin, and Piperacillin + Tazobactum. All the isolates showed a good sensitivity towards Nitrofurantoin (82%). ESBL production was found to be 70.6% in Escherichia coli and 29.4% in Klebsiella pneumonia. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatient UTIs challenging. Ampicillin, Cotrimoxazole and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help in appropriate and judicious antibiotic usage in a health care setup.

Crude Glycerol Affects Canine Sperm Motility: Computer Assisted Semen Analysis in vitro

Target of this study was the analysis of the impact of crude glycerol on canine spermatozoa motility, morphology, viability, and membrane integrity. Experiments were realized in vitro. In the study, semen from 5 large dog breeds was used. They were typical representatives of large breeds, coming from healthy rearing, regularly vaccinated and integrated to the further breeding. Semen collections were realized at the owners of animals and in the veterinary clinic. Subsequently the experiments were realized at the Department of Animal Physiology of the SUA in Nitra. The spermatozoa motility was evaluated using CASA analyzer (SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C for 5 hours. In the study, 13 motility parameters were evaluated. Generally, crude glycerol has generally negative effect on spermatozoa motility. Morphological analysis was realized using Hancock staining and the preparations were evaluated at magnification 1000x using classification tables of morphologically changed spermatozoa. Data clearly detected the highest number of morphologically changed spermatozoa in the experimental groups (know twisted tails, tail torso and tail coiling). For acrosome alterations swelled acrosomes, removed acrosomes and acrosomes with undulated membrane were detected. In this study also the effect of crude glycerol on spermatozoa membrane integrity were analyzed. The highest crude glycerol concentration significantly affects spermatozoa integrity. Results of this study show that crude glycerol has effect of spermatozoa motility, viability, and membrane integrity. Detected changes are related to crude glycerol concentration, temperature, as well as time of incubation.

Identifying Business Incubators Based on Their Real Activities: Evidence from China

Past literature on business incubators distinguished incubators based on their mission statements. However, more and more mission statements become a slogan rather than a reality. It is therefore more appropriate to identify business incubators based on their real activities, rather than the missions they declared. With a sample of technology business incubators (TBIs) in China, we try to investigate business incubators’ real activities by examining the incubation efficiency along the following five dimensions, i.e., survival of new ventures, technology transfer, local economic growth, job creation, and profit generation. Furthermore, we identified six types of business incubators. The results indicate that generally Chinese TBIs have a greater preference for acquiring profits over other dimensions. 

A Piscan Ulcerative Aeromonas Infection

In the immunologic sense, clinical infection is a state of failure of the immune system to combat the pathogenic weapon of the bacteria invading the host. A motile gram negative vibroid organism associated with marked mono and poly nuclear cell responses was traced during the examination of a clinical material from an infected common carp Cyprinus carpio. On primary plate culture, growth was shown to be pure, dense population of an Aeromonas-like colony morphotype. The pure isolate was found to be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and 37C, oxidase positive utilizes glucose through fermentative pathway, resist 0/129 and novobiocin, produces alanine and lysine decarboxylases but non-producing ornithine dehydrolases. Tests for the in vitro determinants of pathogenicity has shown to be; Betahaemolytic onto blood agar, gelatinase, casienase and amylase producer. Three in vivo determinants of pathogenicity were tested as, the lethal dose fifty, the pathogenesis and pathogenicity. It was evident that 0.1 milliliter of the causal bacterial cell suspension of a density 1 x 107 CFU/ml injected intramuscularly into an average of 100gms fish toke five days incubation period, then at the day six morbidity and mortality were initiated. LD50 was recorded at the day 12 post-infection. Use of an LD50 doses to study the pathogenicity, reveals mononuclear and polynuclear cell responses, on examining the stained direct films of the clinical materials from the experimentally infected fish. Re-isolation tests confirm that the reisolant is same. The course of the infection in natural case was shown manifestation of; skin ulceration, haemorrhage and descaling. On evisceration, the internal organs were shown; congestion in the intestines, spleen and, air sacs. The induced infection showed a milder form of these manifestations. The grading of the virulence of this organism was virulent causing chronic course of infections as indicated from the pathogenesis and pathogenicity studies. Thus the infectious bacteria were consistent with Aeromonas hydrophila, and the infection was chronic.

Cissampelos capensis Rhizome Extract Induces Intracellular ROS Production, Capacitation and DNA Fragmentation in Human Spermatozoa

More than 3000 plants of notable phyto-therapeutic value grow in South Africa; these include Cissampelos capensis, commonly known in Afrikaans as dawidjie or dawidjiewortel. C. capensis is the most significant and popular medicinal plant used by the Khoisan as well as other rural groups in the Western region of South Africa. Its rhizomes are traditionally used to treat male fertility problems. Yet, no studies have investigated the effects of this plant or its extracts on human spermatozoa. Therefore, this study aimed at investigating the effects of C. capensis rhizome extract (CRE) fractions on ejaculated human spermatozoa in vitro. Spermatozoa from a total of 77 semen samples were washed with human tubular fluid medium supplemented with bovine serum albumin (HTF-BSA) and incubated for 2 hours with 20 μg/ml progesterone (P4) followed by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200 μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH, F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A sample without addition of CRE fractions served as control. Samples were analyzed for sperm motility, reactive oxygen species (ROS), DNA-fragmentation, acrosome reaction and capacitation. Results showed that F1 resulted in significantly higher values for ROS, capacitation and hyper-activation compared to F2, F3, and F4 with P4-stimulated samples generally having higher values. No significant effect was found for the other parameters. In conclusion, alkaloids present in F1 of CRE appear to have triggered sperm intrinsic ROS production leading to sperm capacitation and acrosome reaction induced by P4.

Mercury Removal Using Pseudomonas putida (ATTC 49128): Effect of Acclimatization Time, Speed and Temperature of Incubator Shaker

Microbes have been used to solve environmental problems for many years. The role of microorganism to sequester, precipitate or alter the oxidation state of various heavy metals has been extensively studied. Treatment using microorganism interacts with toxic metal are very diverse. The purpose of this research is to remove the mercury using Pseudomonas putida (P. putida), pure culture ATTC 49128 at optimum growth parameters such as techniques of culture, acclimatization time and speed of incubator shaker. Thus, in this study, the optimum growth parameters of P. putida were obtained to achieve the maximum of mercury removal. Based on the optimum parameters of P. putida for specific growth rate, the removal of two different mercury concentration, 1 ppm and 4 ppm were studied. From mercury nitrate solution, a mercuryresistant bacterial strain which is able to reduce from ionic mercury to metallic mercury was used to reduce ionic mercury. The overall levels of mercury removal in this study were between 80% and 89%. The information obtained in this study is of fundamental for understanding of the survival of P. putida ATTC 49128 in mercury solution. Thus, microbial mercury removal is a potential bioremediation for wastewater especially in petrochemical industries in Malaysia.

Phyllantus niruri Protects against Fe2+ and SNP Induced Oxidative Damage in Mitochondrial Enriched Fractions of Rats Brain

The potential neuroprotective effect of Phyllantus nuriri against Fe2+ and sodium nitroprusside (SNP) induced oxidative stress in mitochondria of rats brain was evaluated. Cellular viability was assessed by MTT reduction, reactive oxygen species (ROS) generation was measured using the probe 2,7-dichlorofluoresce indiacetate (DCFH-DA). Glutathione content was measured using dithionitrobenzoic acid (DTNB). Fe2+ (10μM) and SNP (5μM) significantly decreased mitochondrial activity, assessed by MTT reduction assay, in a dose-dependent manner, this occurred in parallel with increased glutathione oxidation, ROS production and lipid peroxidation end-products (thiobarbituric acid reactive substances, TBARS). The co-incubation with methanolic extract of Phyllantus nuriri (10-200 μg/ml) reduced the disruption of mitochondrial activity, gluthathione oxidation, ROS production as well as the increase in TBARS levels caused by both Fe2+ and SNP in a dose dependent manner. HPLC analysis of the extract revealed the presence of gallic acid (20.540.01), caffeic acid (7.930.02), rutin (25.310.05), quercetin (31.280.03) and kaemferol (14.360.01). This result suggests that these phytochemicals account for the protective actions of P. niruri against Fe2+ and SNP -induced oxidative stress. Our results show that P. nuriri consist important bioactive molecules in the search for an improved therapy against the deleterious effects of Fe2+, an intrinsic producer of reactive oxygen species (ROS), that leads to neuronal oxidative stress and neurodegeneration.

Selection of Pichia kudriavzevii Strain for the Production of Single-Cell Protein from Cassava Processing Waste

A total of 115 yeast strains isolated from local cassava processing wastes were measured for crude protein content. Among these strains, the strain MSY-2 possessed the highest protein concentration (>3.5 mg protein/mL). By using molecular identification tools, it was identified to be a strain of Pichia kudriavzevii based on similarity of D1/D2 domain of 26S rDNA region. In this study, to optimize the protein production by MSY-2 strain, Response Surface Methodology (RSM) was applied. The tested parameters were the carbon content, nitrogen content, and incubation time. Here, the value of regression coefficient (R2) = 0.7194 could be explained by the model which is high to support the significance of the model. Under the optimal condition, the protein content was produced up to 3.77 g per L of the culture and MSY-2 strain contains 66.8 g protein per 100 g of cell dry weight. These results revealed the plausibility of applying the novel strain of yeast in single-cell protein production.

Morphological Interaction of Porcine Oocyte and Cumulus Cells Study on in vitro Oocyte Maturation Using Electron Microscopy

Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by Zona pellucida with layer of cumulus cells ranging between 59.29-202.14 μm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 μg/mL porcine follicle-stimulating hormone, 1 μg/mL LH, 1μg/mL estradiol with ethanol, and 50 μg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into Zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Application of Statistical Approach for Optimizing CMCase Production by Bacillus tequilensis S28 Strain via Submerged Fermentation Using Wheat Bran as Carbon Source

Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

A β-mannanase from Fusarium oxysporum SS-25 via Solid State Fermentation on Brewer’s Spent Grain: Medium Optimization by Statistical Tools, Kinetic Characterization and Its Applications

This study is concerned with the optimization of fermentation parameters for the hyper production of mannanase from Fusarium oxysporum SS-25 employing two step statistical strategy and kinetic characterization of crude enzyme preparation. The Plackett-Burman design used to screen out the important factors in the culture medium revealed 20% (w/w) wheat bran, 2% (w/w) each of potato peels, soyabean meal and malt extract, 1% tryptone, 0.14% NH4SO4, 0.2% KH2PO4, 0.0002% ZnSO4, 0.0005% FeSO4, 0.01% MnSO4, 0.012% SDS, 0.03% NH4Cl, 0.1% NaNO3 in brewer’s spent grain based medium with 50% moisture content, inoculated with 2.8×107 spores and incubated at 30oC for 6 days to be the main parameters influencing the enzyme production. Of these factors, four variables including soyabean meal, FeSO4, MnSO4 and NaNO3 were chosen to study the interactive effects and their optimum levels in central composite design of response surface methodology with the final mannanase yield of 193 IU/gds. The kinetic characterization revealed the crude enzyme to be active over broader temperature and pH range. This could result in 26.6% reduction in kappa number with 4.93% higher tear index and 1% increase in brightness when used to treat the wheat straw based kraft pulp. The hydrolytic potential of enzyme was also demonstrated on both locust bean gum and guar gum.