Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes

Five agricultural wastes were evaluated in submerged fermentation for xylanolytic enzymes production by Penicillium janthinellum. The wastes were hydrolyzed in acid medium and the liquid fraction was used for cultivation. Corn cob (55.3 U/mL) and oat husk (54.8 U/mL) were the best inducers of xylanase. Sugar cane bagasse (23.0 U/mL) and corn husk (23.8 U/mL) were moderately good, while cassava peel was negligible. Protease production was very low in all agro-industrial residues. The maximum biomass yields were 1.30 and 1.17 g/L for cassava peel and corn husk after 180 h, respectively. Xylanolytic activity showed a cell growth associated profile.     

Polyurethane foam as an inert carrier for the production of L(+)-lactic acid by Lactobacillus casei under solid-state fermentation

AIM: Production of L-lactic acid in solid-state fermentation (SSF) using polyurethane foam (PUF) as inert support moistened with cassava bagasse starch hydrolysate. METHODS AND RESULTS: PUF impregnated with cassava bagasse starch hydrolysate as major carbon source was used for the production of L-lactic acid using Lactobacillus casei in solid-state condition. The key parameters such as reducing sugar, inoculum size and nutrient mixture were optimized by statistical approach using response surface methodology. More than 95% conversion of sugars to lactic acid from 4 g reducing sugar per gram dry support was attained after 72 h when the inert substrate was moistened with 6.5 ml of nutrient solution and inoculated with 1.5 x 10(9) CFU of L. casei. While considering the lactate yield based on the solid support used, a very high yield of 3.88 g lactic acid per gram PUF was achieved. CONCLUSION: PUF acted as an excellent inert support for L. casei and provided a platform for the utilization of starchy waste hydrolysate in a lower reactor volume. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a cost effective cultivation of lactic acid bacteria for producing lactic acid from agro based waste products such as cassava bagasse. This is the first report on the exploitation of PUF as an inert support for lactate production under SSF.     

Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production

In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8?g:10?g:2?g yielded the highest enzyme production of 201.6?U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5?×?10⁶ spores/mL inoculum, which gave the highest enzyme activity of 389.5?U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2?g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300?g raw cassava chips/L with cane molasses.     

木薯粉同步糖化发酵(SSF)产丁二酸

【目的】通过优化产琥珀酸放线杆菌GXAS137同步糖化发酵木薯粉产丁二酸的发酵培养基,提高丁二酸产量,降低生产成本。【方法】在单因素试验的基础上,先利用Plackett-Burman试验设计筛选出影响丁二酸发酵的重要参数,再采用正交试验确定重要参数的最佳水平。【结果】价格低廉玉米浆可用作氮源,影响丁二酸产量的重要参数是木薯粉、玉米浆、碱式碳酸镁和糖化酶浓度。最佳条件为(g/L):木薯粉100,玉米浆14,糖化酶2.0 AGU/g底物,碱式碳酸镁75。优化后丁二酸产量达到69.31 g/L,丁二酸得率为90.01%,生产强度为1.44 g/(L·h)。与初始条件(52.34 g/L)相比,丁二酸浓度提高了32.42%。并利用1.3 L发酵罐对SSF与SHF两种发酵工艺进行了比较,SSF丁二酸产量(72.21 g/L)远高于SHF(56.86 g/L)。【结论】产琥珀酸放线杆菌同步糖化发酵木薯粉丁二酸产量高,生产成本低,具有较好的工业化应用前景。     

Production of multiple extracellular enzyme activities by novel submerged culture of <Emphasis Type="Italic">Aspergillus kawachii</Emphasis> for ethanol production from raw cassava flour

Cassava is a starch-containing root crop that is widely used as a raw material in a variety of industrial applications, most recently in the production of fuel ethanol. In the present study, ethanol production from raw (uncooked) cassava flour by simultaneous saccharification and fermentation (SSF) using a preparation consisting of multiple enzyme activities from Aspergillus kawachii FS005 was investigated. The multi-activity preparation was obtained from a novel submerged fermentation broth of A. kawachii FS005 grown on unmilled crude barley as a carbon source. The preparation was found to consist of glucoamylase, acid-stable α-amylase, acid carboxypeptidase, acid protease, cellulase and xylanase activities, and exhibited glucose and free amino nitrogen (FAN) production rates of 37.7 and 118.7 mg/l/h, respectively, during A. kawachii FS005-mediated saccharification of uncooked raw cassava flour. Ethanol production from 18.2% (w/v) dry uncooked solids of raw cassava flour by SSF with the multi-activity enzyme preparation yielded 9.0% (v/v) of ethanol and 92.3% fermentation efficiency. A feasibility study for ethanol production by SSF with a two-step mash using raw cassava flour and the multi-activity enzyme preparation manufactured on-site was verified on a pilot plant scale. The enzyme preparation obtained from the A. kawachii FS005 culture broth exhibited glucose and FAN production rates of 41.1 and 135.5 mg/l/h, respectively. SSF performed in a mash volume of about 1,612 l containing 20.6% (w/v) dry raw cassava solids and 106 l of on-site manufactured A. kawachii FS005 culture broth yielded 10.3% (v/v) ethanol and a fermentation efficiency of 92.7%.     

Production of pigments byMonascus purpureus using sugar-cane bagasse in roller bottle cultures

Solid-state fermentation, using sugar-cane bagasse, and submerged fermentation, using a semi-synthetic medium, were performed for pigment production byMonascus purpureus in both stationary and rotary conditions. Rotary cultures gave higher yields of crude red and yellow pigments than stationary cultures whereas twice the amount was synthesized at an earlier time (day 8) in liquid medium (1,285U yellow pigment/bottle, 1,728U red pigment/bottle). Supplementing the liquid medium with 0.6% (v/v) corn oil doubled the extracellular pigment yield but halved fungal growth.     

Kinetics of Gibberella fujikuroi growth and gibberellic acid production by solid-state fermentation in a packed-bed column bioreactor

In this work the growth of Gibberella fujikuroi and gibberellic acid (GA3) production were studied using coffee husk and cassava bagasse as substrates in a packed-bed column bioreactor connected to a gas chromatograph for exit gas analysis. With the respirometric data, a logarithmic correlation between accumulated CO2 and biomass production was determined, and the kinetics of the fungal growth was compared for estimated and experimental data. The solid medium consisted of coffee husk (pretreated with alkali solution), mixed with cassava bagasse (7:3 dry weight basis), with a substrate initial pH of 5.2 and moisture of 77%. Cultivation was carried out in glass columns, which were packed with preinoculated substrate and with forced aeration of 0.24 L of air/[h (g of substrate)] for the first 3 days, and 0.72 L of air/[h (g of substrate)] for the remaining period. The maximum specific growth rate (microm) obtained was 0.052 h(-1) (between 24 and 48 h of fermentation). A production of 0.925 g of GA3/kg of substrate was achieved after 6 days of fermentation.     

Optimization of the production of aroma compounds by Kluyveromyces marxianus in solid-state fermentation using factorial design and response surface methodology

Studies were carried out for the production of aroma compounds in solid-state fermentation using factorial design and response surface methodology (RSM) experiments. Five agro-industrial residues were evaluated as substrate for cultivating a strain of Kluyveromyces marxianus. The results proved the feasibility of using cassava bagasse and giant palm bran (Opuntia ficus indica) as substrates to produce fruity aroma compounds by the yeast culture. In order to test the influence of the process parameters on the culture to produce volatile compounds, two statistical experimental designs were performed. The parameters studied were initial substrate pH, addition of glucose, cultivation temperature, initial substrate moisture and inoculum size. Using a 2(5) factorial design, addition of glucose and initial pH of the substrate was found statistically significant for aroma compounds production on palm bran. Although this experimental design showed that addition of glucose did not have a significant role with cassava bagasse, 2(2) factorial design revealed that glucose addition was significant at higher concentrations. Head-space analysis of the culture by gas chromatography showed the production of nine and eleven compounds from palm bran and cassava bagasse, respectively, which included alcohols, esters and aldehyde. In both the cases, two compounds remained unidentified and ethyl acetate, ethanol and acetaldehyde were the major compounds produced. Esters produced were responsible for the fruity aroma in both the cases. With palm bran, ethanol was the compound produced in highest concentration, and with cassava bagasse (both supplemented with 10% glucose), ethyl acetate was produced at highest concentration, accumulating 418 and 1395μmoll(-1) head-spaceg(-1) substrate in 72h, respectively.     

Breeding and growth of Rhizopus in raw cassava by solid state fermentation

Nineteen Rhizopus strains were selected and tested for their growth capacity on raw cassava starch and their ability to produce amylase when grown on solid-state fermentations. Only three strains grew significantly on this natural substrate. Glucoamylase production was higher on raw cassava than on cooked cassava. After 48 h of fermentation, the protein content of cassava was increased from 1.75% to 11.3%. The byproducts of fermentation were fumaric acid, lactid acid and ethanol.     

黑曲霉AF-98固体发酵产纤维素酶的产酶条件研究

通过单因子及正交试验,对黑曲霉AF-98固体发酵产纤维素酶的产酶条件进行了探讨。其优化的产酶条件为:甘蔗渣3g,麸皮2g,加含尿素为0.15%的Mandels营养液25mL(加水比1:5),调初始pH5.0,28℃发酵72h。在此优化条件下,纤维素酶活力可达7.56u/g干曲。     

Statistical optimization of <Emphasis Type="Italic">α</Emphasis>-amylase production by <Emphasis Type="Italic">Bacillus brevis</Emphasis> MTCC 7521 in solid-state fermentation using cassava bagasse

Production of α-amylase under solid-state fermentation by Bacillus brevis MTCC 7521 has been investigated using cassava bagasse as the substrate, one of the major solid wastes released during extraction of starch from cassava (Manihot esculenta). Response surface methodology was used to evaluate the effect of the main variables, i.e. incubation period (36 h), moisture holding capacity (60%), pH (7.0) and temperature (60°C) on enzyme production by applying a full factorial central composite design. The maximum hydrolysis of soluble starch (85%) and cassava starch (75%) was obtained with the application of 4 mL (≈ 14,752 units) of B. brevis crude enzyme after 5 h of incubation.     

Growth of Chaetomium cellulolyticum in solid-state fermentation of sugar cane bagasse treated with water and alkali at several liquid/solid ratios

Sugar cane bagasse was water- or alkali-treated at three liquid/solid (L/S) ratios and its digestibility was measured as microbial protein production of Chaetomium cellulolyticum grown on solid-state fermentation columns. The treatments significantly enhanced fungus growth compared to non-treated bagasse, which was used as a control, although the composition of bagasse did not change greatly. Alkali-treated bagasse reached an average protein content of about 7.6% and the lower the L/S ratio, the higher the protein content. L/S ratio did not have an effect in water-treated bagasse. Protein content of water-treated bagasse was also high, approximately 80% of that one of alkali-treated bagasse. Both treatments look promising to enhance sugar cane bagasse potential as an animal feed.Currently at Universidad Francisco de Miranda. Coro, Venezuela.Currently at Ciclo Básico, Facultad de Ingeniería, Universidad del Zulia. Maracaibo, Venezuela.     

Production of raw cassava starch-degrading enzyme by Penicillium and its use in conversion of raw cassava flour to ethanol

A newly isolated strain Penicillium sp. GXU20 produced a raw starch-degrading enzyme which showed optimum activity towards raw cassava starch at pH 4.5 and 50 °C. Maximum raw cassava starch-degrading enzyme (RCSDE) activity of 20 U/ml was achieved when GXU20 was cultivated under optimized conditions using wheat bran (3.0% w/v) and soybean meal (2.5% w/v) as carbon and nitrogen sources at pH 5.0 and 28 °C. This represented about a sixfold increment as compared with the activity obtained under basal conditions. Starch hydrolysis degree of 95% of raw cassava flour (150 g/l) was achieved after 72 h of digestion by crude RCSDE (30 U/g flour). Ethanol yield reached 53.3 g/l with fermentation efficiency of 92% after 48 h of simultaneous saccharification and fermentation of raw cassava flour at 150 g/l using the RCSDE (30 U/g flour), carried out at pH 4.0 and 40 °C. This strain and its RCSDE have potential applications in processing of raw cassava starch to ethanol.     

Process design and optimization of bioethanol production from cassava bagasse using statistical design and genetic algorithm

Abstract

Bioethanol production from agro-industrial residues is gaining attention because of the limited production of starch grains and sugarcane, and food–fuel conflict. The aim of the present study is to maximize the bioethanol production using cassava bagasse as a feedstock. Enzymatic liquefaction, by α-amylase, followed by simultaneous saccharification and fermentation (SSF), using glucoamylase and Zymomonas mobilis MTCC 2427, was investigated for bioethanol production from cassava bagasse. The factors influencing ethanol production process were identified and screened for significant factors using Plackett–Burman design. The significant factors (cassava bagasse concentration (10–50?g/L), concentration of α-amylase (5–25% (v/v), and temperature of fermentation (27–37?°C)) were optimized by employing Box–Behnken design and genetic algorithm. The maximum ethanol concentrations of 25.594?g/L and 25.910?g/L were obtained from Box–Behnken design and genetic algorithm, respectively, under optimum conditions. Thus, the study provides valuable insights in utilizing the cost-effective industrial residue, cassava bagasse, for the bioethanol production.     

固态发酵木薯酒精渣生产生物饲料菌种的筛选

目的:筛选获得能混合固态发酵木薯酒精渣生产生物饲料的真菌组合.方法:利用木薯酒精渣堵养基,初筛能在其上良好生长的植物内生真菌菌株,再将这些菌株两两组合进行固态混菌发酵、添加酵母混菌发酵,测定产物中粗蛋白和粗纤维的含量,获得能有效降低木薯酒精渣中粗纤维、提高粗蛋白含量的菌株组合.结果:菌株G4与C15、Q4与C32混菌发酵效果最好,可将粗蛋白质含最从底物的1.42%分别提高到产物的16.08%与18.54%(于基),粗纤维含量从底物的32.41%降低到27.57%与26.59%.添加酵母培养后,两个组合产物中粗蛋白质含量可进一步提高到21.79%与23.56%,而粗纤维含量几乎无变化.结论:菌株G4(黑曲霉)、C15(白地霉)与郎比可假丝酵母,Q4(黑曲霉)、C32(青霉)与季也蒙假丝酵母可用作混菌固态发酵木薯酒精渣生产生物饲料的菌种.     

Optimization of bagasse,nutrients and initial moisture ratios on the yield of penicillin in solid-state fermentation

An advanced solid-state fermentation (SSF) system (liquid medium absorbed on an inert support) has been applied to antibiotic production. The main components of this solid medium are: support (sugarcane bagasse), nutrients and water. The first two are solids and have to be considered to calculate the initial moisture content (IMC) of the medium. Earlier work indicated the importance of using high IMCs and concentrated media to obtain high penicillin yields in SSF. Nevertheless, the present work shows that high values of IMC or nutrients content can stimulate or inhibit penicillin production, depending on the strategy used to compensate this change (i.e. the proportions of the other two components). Conversely, increasing bagasse content always showed an inhibitory effect on the production. Since penicillin production depends on the combinations of these components, a global approach was used. The effect of the proportions of the three components on penicillin production was studied by means of a triangle of combinations and a 3D graph. It was possible to establish that high penicillin production is only obtained in a zone of low support content (10–12.5%). Surprisingly, one production maximum was observed in a zone of low moisture and high nutrients content (62 and 25.5% respectively); and another one in a zone of high moisture and a relatively low nutrients concentration (75.5 and 12.4% respectively).     

Potential of solid state fermentation for production of L(+)-lactic acid by Rhizopus oryzae

Production of l(+)-lactic acid by Rhizopus oryzae NRRL 395 was studied in solid medium on sugar-cane bagasse impregnated with a nutrient solution containing glucose and CaCO₃. A comparative study was undertaken in submerged and solid-state cultures. The optimal concentrations in glucose were 120 g/l in liquid culture and 180 g/l in solid-state fermentation corresponding to production of l(+)-lactic acid of 93.8 and 137.0 g/l, respectively. The productivity was 1.38 g/l per hour in liquid medium and 1.43 g/l per hour in solid medium. However, the fermentation yield was about 77% whatever the medium. These figures are significant for l(+)-lactic acid production.     

木聚糖酶生产菌株的筛选及产酶条件的优化

以甘蔗渣半纤维素为碳源,从垃圾场土壤中分离到6株分解半纤维素的菌株。通过固态发酵的木聚糖酶活力比较筛选到1株木聚糖酶活力较高的菌株。该菌株18S rDNA序列与曲霉（Aspergillus sp.）的同源性达97%,根据对菌株形态学分析和18S rDNA序列分析的结果,将该菌株鉴定为曲霉HQ3。HQ3的最佳产酶条件为：甘蔗渣:麸皮为7:3（W/W）,固液比为1:4（W/W）,尿素0.4 %,pH7.0,温度30℃,发酵产酶时间4 d。在最佳产酶条件下,其木聚糖酶活最高可达3421U/g干曲。     

Production of Xylanase by Thermophilic Melanocarpus albomyces IIS-68

Melanocarpus albomyces IIS-68, a thermophilic fungus was used for the production of extracellular xylanase on various agroresidues in solid-state fermentation (SSF). Growth on untreated wheat straw and sugar cane bagasse supported xylanase production, while rice straw and rice husk did not. Alkali treatment and acid chlorite treatment of these latter substrates, which lead to extensive delignification, enhanced xylanase production. In contrast, these treatments caused a decline in xylanase activity on wheat straw and bagasse. Acetyl esterase was produced concurrently with xylanase, maximal activity being produced on bagasse. Enzyme production was higher in SSF than in submerged fermentation (SmF). Studies with electron micrographs indicated that culture filtrate proteins were able to degrade wall polymers.     

Production and isolation of chitosan by submerged and solid-state fermentation from Lentinus edodes

A method for the laboratory-scale production and isolation of chitosan (polyglucosamine) by liquid and solidstate fermentation from Lentinus edodes was developed. The yields of isolated chitosan were 120 mg/L of fermentation medium under liquid fermentation conditions and 6.18 g/kg of fermentation medium under solid-state fermentation conditions. The latter method, which gives up to 50 times yields than other chitosan production methods from fungi, provides a new flexible and easily scaledup procedure for the production of low acetylation degree chitosan. (c) 1996 John Wiley & Sons, Inc.