Solid-State Bioconversion of Pineapple Residues into Kojic Acid by Aspergillus flavus: A Prospective Study

Kojic acid is an organic acid that is widely used as an ingredient for dermatological products, precursor for flavor enhancer and also as anti-inflammatory drug. The present study was undertaken to test the feasibility of pineapple residues as substrate for kojic acid production by Aspergillus flavus Link 44-1 via solid-state fermentation. The effect of initial moisture content, pH and incubation time on kojic acid fermentation was investigated. The best initial moisture content for kojic acid production from pineapple residues was observed at 70% (v/w) whereas initial culture pH 2.5 was identified to give high production of kojic acid. The optimal range of incubation time was identified between 8 and 14 days of incubation which corresponded to highest range of kojic acid produced. The results from this study pronounce the promising usability of pineapple residues as alternative substrate for kojic acid production by A. flavus Link 44-1.

Microbial Oil Production by Isolated Oleaginous Yeast Torulaspora globosa YU5/2

Microbial oil was produced by soil isolated oleaginous yeast YU5/2 in flask-batch fermentation. The yeast was identified by molecular genetics technique based on sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA and it was identified as Torulaspora globosa. T. globosa YU5/2 supported maximum values of 0.520 g/L/d, 0.472 g lipid/g cells, 4.16 g/L, and 0.156 g/L/d for volumetric lipid production rate, and specific yield of lipid, lipid concentration, and specific rate of lipid production respectively, when culture was performed in nitrogen-limiting medium supplemented with 80g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sweet potato tubers hydrolysates as carbon source.

Simultaneous Saccharification and Fermentation(SSF) of Sugarcane Bagasse - Kinetics and Modeling

Simultaneous Saccharification and Fermentation (SSF) of sugarcane bagasse by cellulase and Pachysolen tannophilus MTCC *1077 were investigated in the present study. Important process variables for ethanol production form pretreated bagasse were optimized using Response Surface Methodology (RSM) based on central composite design (CCD) experiments. A 23 five level CCD experiments with central and axial points was used to develop a statistical model for the optimization of process variables such as incubation temperature (25–45°) X1, pH (5.0–7.0) X2 and fermentation time (24–120 h) X3. Data obtained from RSM on ethanol production were subjected to the analysis of variance (ANOVA) and analyzed using a second order polynomial equation and contour plots were used to study the interactions among three relevant variables of the fermentation process. The fermentation experiments were carried out using an online monitored modular fermenter 2L capacity. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (32°C), pH (5.6) and fermentation time (110 h). Maximum ethanol concentration (3.36 g/l) was obtained from 50 g/l pretreated sugarcane bagasse at the optimized process conditions in aerobic batch fermentation. Kinetic models such as Monod, Modified Logistic model, Modified Logistic incorporated Leudeking – Piret model and Modified Logistic incorporated Modified Leudeking – Piret model have been evaluated and the constants were predicted.

Wheat Bran Carbohydrates as Substrate for Bifidobacterium lactis Development

The present study addresses problems and solutions related to new functional food production. Wheat (Triticum aestivum L) bran obtained from industrial mill company “Dobeles dzirnavieks”, was used to investigate them as raw material like nutrients for Bifidobacterium lactis Bb-12. Enzymatic hydrolysis of wheat bran starch was carried out by α-amylase from Bacillus amyloliquefaciens (Sigma Aldrich). The Viscozyme L purchased from (Sigma Aldrich) were used for reducing released sugar. Bifidibacterium lactis Bb-12 purchased from (Probio-Tec® CHR Hansen) was cultivated in enzymatically hydrolysed wheat bran mash. All procedures ensured the number of active Bifidobacterium lactis Bb-12 in the final product reached 105 CFUg-1. After enzymatic and bacterial fermentations sample were freeze dried for analysis of chemical compounds. All experiments were performed at Faculty of Food Technology of Latvia University of Agriculture in January- March 2013. The obtained results show that both types of wheat bran (enzymatically treated and non-treated) influenced the fermentative activity and number of Bifidibacterium lactis Bb-12 viable in wheat bran mash. Amount of acidity strongly increase during the wheat bran mash fermentation. The main objective of this work was to create low-energy functional enzymatically and bacterially treated food from wheat bran using enzymatic hydrolysis of carbohydrates and following cultivation of Bifidobacterium lactis Bb-12.

Potential of Agro-Waste Extracts as Supplements for the Continuous Bioremediation of Free Cyanide Contaminated Wastewater

Different agricultural waste peels were assessed for their suitability to be used as primary substrates for the bioremediation of free cyanide (CN-) by a cyanide-degrading fungus Aspergillus awamori isolated from cyanide containing wastewater. The bioremediated CN- concentration were in the range of 36 to 110 mg CN-/L, with Orange (C. sinensis) > Carrot (D. carota) > Onion (A. cepa) > Apple (M. pumila), being chosen as suitable substrates for large scale CN- degradation processes due to: 1) the high concentration of bioremediated CN-, 2) total reduced sugars released into solution to sustain the biocatalyst, and 3) minimal residual NH4- N concentration after fermentation. The bioremediation rate constants (k) were 0.017h-1 (0h < t < 24h), with improved bioremediation rates (0.02189h-1) observed after 24h. The averaged nitrilase activity was ~10 U/L.

Enzymatic Saccharification of Dilute Alkaline Pre-treated Microalgal (Tetraselmis suecica) Biomass for Biobutanol Production

Enzymatic saccharification of biomass for reducing sugar production is one of the crucial processes in biofuel production through biochemical conversion. In this study, enzymatic saccharification of dilute potassium hydroxide (KOH) pre-treated Tetraselmis suecica biomass was carried out by using cellulase enzyme obtained from Trichoderma longibrachiatum. Initially, the pre-treatment conditions were optimised by changing alkali reagent concentration, retention time for reaction, and temperature. The T. suecica biomass after pre-treatment was also characterized using Fourier Transform Infrared Spectra and Scanning Electron Microscope. These analyses revealed that the functional group such as acetyl and hydroxyl groups, structure and surface of T. suecica biomass were changed through pre-treatment, which is favourable for enzymatic saccharification process. Comparison of enzymatic saccharification of untreated and pre-treated microalgal biomass indicated that higher level of reducing sugar can be obtained from pre-treated T. suecica. Enzymatic saccharification of pre-treated T. suecica biomass was optimised by changing temperature, pH, and enzyme concentration to solid ratio ([E]/[S]). Highest conversion of carbohydrate into reducing sugar of 95% amounted to reducing sugar yield of 20 (wt%) from pre-treated T. suecica was obtained from saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1 after 72 h of incubation. Hydrolysate obtained from enzymatic saccharification of pretreated T. suecica biomass was further fermented into biobutanol using Clostridium saccharoperbutyliticum as biocatalyst. The results from this study demonstrate a positive prospect of application of dilute alkaline pre-treatment to enhance enzymatic saccharification and biobutanol production from microalgal biomass.

Optimization of Conditions for Xanthan Gum Production from Waste Date in Submerged Fermantation

Xanthan gum is one of the major commercial biopolymers. Due to its excellent rheological properties xanthan gum is used in many applications, mainly in food industry. Commercial production of xanthan gum uses glucose as the carbon substrate; consequently the price of xanthan production is high. One of the ways to decrease xanthan price, is using cheaper substrate like agricultural wastes. Iran is one of the biggest date producer countries. However approximately 50% of date production is wasted annually. The goal of this study is to produce xanthan gum from waste date using Xanthomonas campestris PTCC1473 by submerged fermentation. In this study the effect of three variables including phosphor and nitrogen amount and agitation rate in three levels using response surface methodology (RSM) has been studied. Results achieved from statistical analysis Design Expert 7.0.0 software showed that xanthan increased with increasing level of phosphor. Low level of nitrogen leaded to higher xanthan production. Xanthan amount, increasing agitation had positive influence. The statistical model identified the optimum conditions nitrogen amount=3.15g/l, phosphor amount=5.03 g/l and agitation=394.8 rpm for xanthan. To model validation, experiments in optimum conditions for xanthan gum were carried out. The mean of result for xanthan was 6.72±0.26. The result was closed to the predicted value by using RSM.

Biogas Yield Potential Research of Tithonia diversifolia in Mesophilic Anaerobic Fermentation in China

BioEnergy is an archetypal appropriate technology and alternate source of energy in rural areas of China, and can meet the basic need for cooking fuel in rural areas. The paper introduces with an alternate mean of research that can accelerate the biogas energy production. Tithonia diversifolia or the Tree marigold can be hailed as mesophillic anaerobic digestion to increase the production of more Bioenergy. Tithonia diversifolia is very native to Mexico and Central America, which can be served as ornamental plants- green manure and can prevent soil erosion. Tithonia diversifolia is widely grown and known to Asia, Africa, America and Australia as well. Nowadays, Considering China’s geographical condition it is found that Tithonia diversifolia is widely growing plant in the many tropical and subtropical regions of southern Yunnan- which can have great usage in accelerating and increasing the Bioenergy production technology. The paper discussed aiming at proving possibility that Tithonia diversifolia can be applied in biogas fermentation and its biogas production potential, the research carried experiment on Tithonia diversifolia biogas fermentation under the mesophilic condition (35 Celsius Degree). The result revealed that Tithonia diversifolia can be used as biogas fermentative material, and 6% concentration can get the best biogas production, with the TS biogas production rate 656mL/g and VS biogas production rate 801mL/g. It is well addressed that Tithonia diversifolia grows wildly in 53 Counties and 9 cities of Yunnan Province, which mainly grows in form of the road side plants, the edge of the field, countryside, forest edge, open space; of which demersum-natures can form dense monospecific beds -causing serious harm to agricultural production landforms threatening the ecological system as a potentially harmful exotic plant. There are also found the three types of invasive daisy alien plants -Eupatorium adenophorum, Eupatorium Odorata and Tithonia diversifolia in Yunnan Province of China-among them the Tithonia diversifolia is responsible for causing serious harm to agricultural production. In this paper we have designed the experimental explanation of Biogas energy production that requires anaerobic environment and some microbes; Tithonia diversifolia plant has been taken into consideration while carrying experiments and with successful resulting of generating more BioEnergy emphasizing on the practical applications of Tithonia diversifolia. This paper aims at- to find a new mechanism to provide a more scientific basis for the development of this plant herbicides in Biogas energy and to improve the utilization throughout the world as well.

Glutamic Acid Production from Potato by Brevibacterium linens

In this study, the possibility of using potato as a substrate for glutamic acid production by Brevibacterium linens was investigated. For preparation of fermentation medium, potato was hydrolyzed by hydrochloridric acid. The medium contained potato hydrolysate, tween 80, mineral solution, glucose, and potassium hydrogen phosphate. The initial pH of the medium was adjusted to 7-7.5. For achieving the optimum time with maximum yield, the beakers containing the medium and the inoculums were incubated in a rotary water bath flask shaker for one to five days. Thin layer choromatography was used for quantitative and qualitative assay of the glutamic acid produced. The results revealed that as fermentation time increased, pH of the fermentation medium significantly decreased (P

Bioconversion of Biodiesel Derived Crude Glycerol by Immobilized Clostridium pasteurianum: Effect of Temperature

Batch fermentation of 5, 10 and 25 g/L biodiesel derived crude glycerol was carried out at 30, 37 and 450C by Clostridium pasteurianum cells immobilized on silica. Maximum yield of 1,3-propanediol (PDO) (0.60 mol/mol), and ethanol (0.26 mol/mol) were obtained from 10 g/L crude glycerol at 30 and 370C respectively. Maximum yield of butanol (0.28 mol/mol substrate added) was obtained at 370C with 25 g/L substrate. None of the three products were detected at 45oC even after 10 days of fermentation. Only traces of ethanol (0.01 mol/mol) were detected at 450C with 5 g/L substrate. The results obtained for 25 g/L substrate utilization were fitted in first order rate equation to obtain the values of rate constant at three different temperatures for bioconversion of glycerol. First order rate constants for bioconversion of glycerol at 30, 37 and 45oC were found to be 0.198, 0.294 and 0.029/day respectively. Activation energy (Ea) for crude glycerol bioconversion was calculated to be 57.62 kcal/mol.

Isolation and Identification of an Acetobacter Strain from Iranian White-Red Cherry with High Acetic Acid Productivity as a Potential Strain for Cherry Vinegar Production in Foodand Agriculture Biotechnology

According to FDA (Food and Drug Administration of the United States), vinegar is definedas a sour liquid containing at least 4 grams acetic acid in 100 cubic centimeter (4% solution of acetic acid) of solution that is produced from sugary materials by alcoholic fermentation. In the base of microbial starters, vinegars could be contained of more than 50 types of volatile and aromatic substances that responsible for their sweet taste and smelling. Recently the vinegar industry has a great proportion in agriculture, food and microbial biotechnology. The acetic acid bacteria are from the family Acetobacteraceae. Regarding to the latest version of Bergy-s Mannual of Systematic Bacteriology that has categorized bacteria in the base of their 16s RNA differences, the most important acetic acid genera are included Acetobacter (genus I), Gluconacetobacter (genus VIII) and Gluconobacter (genus IX). The genus Acetobacter that is primarily used in vinegar manufacturing plants is a gram negative, obligate aerobe coccus or rod shaped bacterium with the size 0.6 - 0.8 X 1.0 - 4.0 μm, nonmotile or motile with peritrichous flagella and catalase positive – oxidase negative biochemically. Some strains are overoxidizer that could convert acetic acid to carbon dioxide and water.In this research one Acetobacter native strain with high acetic acid productivity was isolated from Iranian white – red cherry. We used two specific culture media include Carr medium [yeast extract, 3%; ethanol, 2% (v/v); bromocresol green, 0.002%; agar, 2% and distilled water, 1000 ml], Frateur medium [yeast extract, 10 g/l; CaCO3, 20 g/l; ethanol, 20 g/l; agar, 20 g/l and distilled water, 1000 ml] and an industrial culture medium. In addition to high acetic acid production and high growth rate, this strain had a good tolerance against ethanol concentration that was examined using modified Carr media with 5%, 7% and 9% ethanol concentrations. While the industrial strains of acetic acid bacteria grow in the thermal range of 28 – 30 °C, this strain was adapted for growth in 34 – 36 °C after 96 hours incubation period. These dramatic characteristics suggest a potential biotechnological strain in production of cherry vinegar with a sweet smell and different nutritional properties in comparison to recent vinegar types. The lack of growth after 24, 48 and 72 hours incubation at 34 – 36 °C and the growth after 96 hours indicates a good and fast thermal flexibility of this strain as a significant characteristic of biotechnological and industrial strains.

Quality Properties of Fermented Mugworts and Rapid Pattern Analysis of Their Volatile Flavor Components by Electric Nose Based On SAW (Surface Acoustic Wave) Sensor in GC System

The changes in quality properties and nutritional components in two fermented mugworts (Artemisia capillaries Thumberg, Artemisiaeasiaticae Nakai) were characterized followed by the rapid pattern analysis of volatile flavor compounds by Electric Nose based on SAW(Surface Acoustic Wave) sensor in GC system. There were remarkable decreases in the pH and small changes in the total soluble solids after fermentation. The L (lightness) and b (yellowness) values in Hunter's color system were shown to be decreased, whilst the a (redness) value was increased by fermentation. The HPLC analysis demonstrated that total amino acids were increased in quantity and the essential amino acids were contained higher in A. asiaticaeNakai than in A. capillaries Thumberg. While the total polyphenol contents were not affected by fermentation, the total sugar contents were dramatically decreased. Scopoletinwere highly abundant in A. capillarisThumberg, however, it was not detected in A. asiaticaeNakai. Volatile flavor compounds by Electric Nose showed that the intensity of several peaks were increased much and seven additional flavor peaks were newly produced after fermentation. The flavor differences of two mugworts were clearly distinguished from the image patterns of VaporPrintTM which indicate that the fermentation enables the two mugworts to have subtle flavor differences.

Cyanide and Heavy Metal Concentration of Fermented Cassava Flour (Lafun) Available in the Markets of Ogun and Oyo States of Nigeria

Fermented cassava flours (lafun) sold in Ogun and Oyo States of Nigeria were collected from 10 markets for a period of two months and analysed to determine their safety status. The presence of trace metals was due to high vehicular movement around the drying sites and markets. Cyanide and moisture contents of samples were also determined to assess the adequacy of fermentation and drying. The result showed that sample OWO was found to have the highest amount of 16.02±0.12mg/kg cyanide while the lowest was found in sample OJO with 10.51±0.10mg/kg. The results also indicated that sample TVE had the highest moisture content of 18.50±0.20% while sample OWO had the lowest amount of 12.46±0.47%. Copper and lead levels were found to be highest in TVE with values 28.10mg/kg and 1.1mg/kg respectively, while sample BTS had the lowest values of 20.6mg/kg and 0.05mg/kg respectively. High value of cyanide indicated inadequate fermentation.

Microalgal Lipid Production by Microalgae Chlorella sp. KKU-S2

The objective of this work is to produce heterotrophic microalgal lipid in flask-batch fermentation. Chlorella sp. KKU-S2 supported maximum values of 0.374 g/L/d, 0.478 g lipid/g cells, and 0.112 g/L/d for volumetric lipid production rate, and specific yield of lipid, and specific rate of lipid production, respectively when culture was performed on BG-11 medium supplemented with 50g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sugarcane molasses as carbon source. The main components of fatty acid from extracted lipid were palmitic acid, stearic acid, oleic acid and linoleic acid which similar to vegetable oils and suitable for biodiesel production.

Optimization of Ethanol Fermentation from Pineapple Peel Extract Using Response Surface Methodology (RSM)

Ethanol has been known for a long time, being perhaps the oldest product obtained through traditional biotechnology fermentation. Agriculture waste as substrate in fermentation is vastly discussed as alternative to replace edible food and utilization of organic material. Pineapple peel, highly potential source as substrate is a by-product of the pineapple processing industry. Bio-ethanol from pineapple (Ananas comosus) peel extract was carried out by controlling fermentation without any treatment. Saccharomyces ellipsoides was used as inoculum in this fermentation process as it is naturally found at the pineapple skin. In this study, the capability of Response Surface Methodology (RSM) for optimization of ethanol production from pineapple peel extract using Saccharomyces ellipsoideus in batch fermentation process was investigated. Effect of five test variables in a defined range of inoculum concentration 6- 14% (v/v), pH (4.0-6.0), sugar concentration (14-22°Brix), temperature (24-32°C) and time of incubation (30-54 hrs) on the ethanol production were evaluated. Data obtained from experiment were analyzed with RSM of MINITAB Software (Version 15) whereby optimum ethanol concentration of 8.637% (v/v) was determined. The optimum condition of 14% (v/v) inoculum concentration, pH 6, 22°Brix, 26°C and 30hours of incubation. The significant regression equation or model at the 5% level with correlation value of 99.96% was also obtained.

Screening of Process Variables for the Production of Extracellular Lipase from Palm Oil by Trichoderma Viride using Plackett-Burman Design

Plackett-Burman statistical screening of media constituents and operational conditions for extracellular lipase production from isolate Trichoderma viride has been carried out in submerged fermentation. This statistical design is used in the early stages of experimentation to screen out unimportant factors from a large number of possible factors. This design involves screening of up to 'n-1' variables in just 'n' number of experiments. Regression coefficients and t-values were calculated by subjecting the experimental data to statistical analysis using Minitab version 15. The effects of nine process variables were studied in twelve experimental trials. Maximum lipase activity of 7.83 μmol /ml /min was obtained in the 6th trail. Pareto chart illustrates the order of significance of the variables affecting the lipase production. The present study concludes that the most significant variables affecting lipase production were found to be palm oil, yeast extract, K2HPO4, MgSO4 and CaCl2.

Kinetic and Optimization Studies on Ethanol Production from Corn Flour

Studies on Simultaneous Saccharification and Fermentation (SSF) of corn flour, a major agricultural product as the substrate using starch digesting glucoamylase enzyme derived from Aspergillus niger and non starch digesting and sugar fermenting Saccharomyces cerevisiae in a batch fermentation. Experiments based on Central Composite Design (CCD) were conducted to study the effect of substrate concentration, pH, temperature, enzyme concentration on Ethanol Concentration and the above parameters were optimized using Response Surface Methodology (RSM). The optimum values of substrate concentration, pH, temperature and enzyme concentration were found to be 160 g/l, 5.5, 30°C and 50 IU respectively. The effect of inoculums age on ethanol concentration was also investigated. The corn flour solution equivalent to 16% initial starch concentration gave the highest ethanol concentration of 63.04 g/l after 48 h of fermentation at optimum conditions of pH and temperature. Monod model and Logistic model were used for growth kinetics and Leudeking – Piret model was used for product formation kinetics.

Integrated Cultivation Technique for Microbial Lipid Production by Photosynthetic Microalgae and Locally Oleaginous Yeast

The objective of this research is to study of microbial lipid production by locally photosynthetic microalgae and oleaginous yeast via integrated cultivation technique using CO2 emissions from yeast fermentation. A maximum specific growth rate of Chlorella sp. KKU-S2 of 0.284 (1/d) was obtained under an integrated cultivation and a maximum lipid yield of 1.339g/L was found after cultivation for 5 days, while 0.969g/L of lipid yield was obtained after day 6 of cultivation time by using CO2 from air. A high value of volumetric lipid production rate (QP, 0.223 g/L/d), specific product yield (YP/X, 0.194), volumetric cell mass production rate (QX, 1.153 g/L/d) were found by using ambient air CO2 coupled with CO2 emissions from yeast fermentation. Overall lipid yield of 8.33 g/L was obtained (1.339 g/L of Chlorella sp. KKU-S2 and 7.06g/L of T. maleeae Y30) while low lipid yield of 0.969g/L was found using non-integrated cultivation technique. To our knowledge this is the unique report about the lipid production from locally microalgae Chlorella sp. KKU-S2 and yeast T. maleeae Y30 in an integrated technique to improve the biomass and lipid yield by using CO2 emissions from yeast fermentation.

Cell Growth and Metabolites Produced by Fluorescent Pseudomonad R62 in Modified Chemically Defined Medium

Chemically defined Schlegel-s medium was modified to improve production of cell growth and other metabolites that are produced by fluorescent pseudomonad R62 strain. The modified medium does not require pH control as pH changes are kept within ± 0.2 units of the initial pH 7.1 during fermentation. The siderophore production was optimized for the fluorescent pseudomonad strain in the modified medium containing 1% glycerol as a major carbon source supplemented with 0.05% succinic acid and 0.5% Ltryptophan. Indole-3 acetic acid (IAA) production was higher when L-tryptophan was used at 0.5%. The 2,4- diacetylphloroglucinol (DAPG) was higher with amended three trace elements in medium. The optimized medium produced 2.28 g/l of dry cell mass and 900 mg/l of siderophore at the end of 36 h cultivation, while the production levels of IAA and DAPG were 65 mg/l and 81 mg/l respectively at the end of 48 h cultivation.