Optimization of Solid-State Fermentation with Lactobacillus brevis and Aspergillus oryzae for Trypsin Inhibitor Degradation in Soybean Meal

The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation （SSF） with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology （RSM） with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were： pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were： substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically （57.1 and 89.2% respectively）. L. brevis fermentation did not affect phytic acid （0.4%） and crude fat （5.2%） considerably, whereas A. oryzae fermentation degraded phytic acid （34.8%） and crude fat （22.0%） contents to a certain extent. Crude protein content was increased after both fermentation （6.4 and 12.9% for L. brevis and A. oryzae respectively）. Urease activity was reduced greatly （83.3 and 58.3% for L. brevis and A. oryzae respectively）. In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal.

Spore production in the solid-state fermentation of stevia residue by Trichoderma guizhouense and its effects on corn growth

Trichoderma is an important and widely used plant growth-promoting fungus(PGPF).In this study,stevia residue amended with amino acids hydrolyzed from animal carcasses was used for the production of Trichoderma guizhouense NJAU 4742 by solid-state fermentation,and then its potential to promote corn plant growth was evaluated in combination with chemical fertilizer(CF)or organic fertilizer(OF).The highest spore number of 7×10^(9) CFU g^(–1) fresh weight was obtained under the following optimal parameters:material ratio of 50%(stevia residue:rice bran=1:1),pH value of 3.0(amended with 6.67%amino acids),initial moisture content of 60%,inoculum size of 10%,material thickness of 3 cm and an incubation time of 4 days.The aboveground corn plant biomass obtained with T.guizhouense applied alone and with CF treatments were slightly higher than those of no fertilizer control and CF treatments,respectively.However,T.guizhouense applied with OF significantly(P<0.05)increased aboveground biomass compared to OF and yielded the highest aboveground biomass among all the treatments.Moreover,T.guizhouense applications primarily influenced the fungal bulk soil community composition,among which three OTUs(OTU_(2) and OTU_(9) classified as Chaetomium,and OTU_(4)classified as Trichoderma)were stimulated in both bulk and rhizosphere soil.Notably,a specific OTU_(3)(Phymatotrichopsis)was only stimulated by T.guizhouense applied with OF,possibly leading to high soil productivity.These results show that it is feasible to employ stevia residue in the eco-friendly fermentation of T.guizhouense,which is strongly suggested for enhancing OF applications.

The variation of two extracellular enzymes and soybean meal bitterness during solid-state fermentation of Bacillus subtilis

The debittering effect of extracellular enzymes from Bacillus subtilis ACCC 01746 was studied using soybean meal as a substrate for solid-state fermentation(SSF).Results showed that B.subtilis produces proteases and carboxypeptidase in the early stage of SSF(0–8 h).Proteases are dominant and can hydrolyze the soybean protein into long-chain peptides with mild bitterness.Carboxypeptidase production is dominant at 8–16 h SSF,at which point soybean protein is further hydrolyzed and bitterness is enhanced.The strain then produces additional carboxypeptidase after 16 h,and bitterness is reduced.We compared the amino acid composition of the hydrolysates from soybean protein isolates to that of the fermented liquid of SSF.In the hydrolysates from soybean protein isolates that exhibit strong bitterness,62.81%of amino acids are hydrophobic and occur in the form of peptides.In the fermented liquid from soybean meal,16.22%of amino acids are hydrophobic and are mainly present in the form of free amino acids.The bitterness of fermented soybean hydrolysate is reduced from 5 to 0 when fermented for 24 h,suggesting that B.subtilis can effectively reduce bitterness,possibly due to the carboxypeptidase.Enzyme analysis shows that B.subtilis excretes carboxypeptidase during growth.The amino acids phenylalanine,alanine,tyrosine,and leucine at the C-terminal of the soybean bitter peptides in hydrolysates are cleaved in the presence of carboxypeptidase,resulting in complete debitterness.

Immobilization of trophic anaerobic acetogen for semi-continuous syngas fermentation

The massive consumption of fossil energy force s people to find new source s of energy.Syngas fermentation has become a hot research field as its high potential in renewable energy production and sustainable development.In this study,trophic anaerobic acetogen Morella thermoacetica was successfully immobilized by calcium alginate embedding method.The ability of the immobilized cells on production of acetic acid through syngas fermentation was compared in both airlift and bubble column bioreactors.The bubble column bioreactor was selected as the better type of bioreactor.The production of acetic acid reached 32.3 g·L^(-1) in bubble column bioreactor with a space-time yield of 2.13 g·L^(-1)·d^(-1).The immobilized acetogen could be efficiently reused without significant lag period,even if exposed to air for a short time.A semi-continuous syngas fermentation was performed using immobilized cells,with an average space-time acetic acid yield of 3.20 g·L^(-1)·d^(-1).After 30 days of fermentation,no significant decrea se of the acetic acid production rate was observed.

Effects of Application of Cassava Alcohol Anaerobic Fermentation Liquid and Mulching of Film on Overwintering and Next Year's Growth and Yield of Banana

The effects of application of cassava alcohol anaerobic fermentation liquid and mulching of film on overwintering and next year's growth and yield of banana were investigated in this study.The results showed that the application of cassava alcohol anaerobic fermentation liquid with COD concentration of 1 200-2 000 mg/L(900 m^3/hm^2) and mulching of film significantly increased the soil organic matter and available potassium contents in banana orchard,improved the cold resistance of banana seedlings,increased the pseudostem height,pseudostem circumference,green leaf number and chlorophyll content of next year's banana seedlings,brought the flower bud emergence period and harvest period forward,and significantly increased the yield per plant.

Consideration and development of anaerobic fermentation equipment on solid organic rejectamenta

This article, based on investigation and analysis, existed anaerobic fermentation equipment, with a view to the production of high-latitude area, pointed out the thought of exploiting efficient anaerobic fermentation equipment, including： the settting independent equipment of acidogenic phase and methanogenic phase; the applying conbination of AF and UASB in methanogenic phase; adopting efficient sludge inverse flowing equipment and the technique of flora enrichment, and efficient method of saving energy and thermal retardation; adopting autocontrol which could make the equipment run efficiently and stably.

Novel Protease from Aspergillus tamarii URM4634: Production and Characterization Using Inexpensive Agroindustrial Substrates by Solid-State Fermentation

This study reports the protease production from Aspergillus tamarii using agroindustrial residues as substrate for solid-state fermentation (SSF) and biochemical characterization. The highest protease production was obtained using wheat bran as substrate at 72 h fermentation with maximum proteolytic activity of 401.42 U/mL, collagenase of 243.0 U/mL and keratinase of 19.1 U/mL. The protease exhibited KM = 18.7 mg/mL and Vmax = 28.5 mg/mL/min. The optimal pH was 8.0 and stable in a wide pH range (5.0 - 11.0) during 24 h. The optimum temperature was 40°C. The proteolytic activity was inhibited by Cu2+ (33.98%) and Hg2+ (22.69%). The enzyme was also inhibited by PMSF (65.11%), indicating that is a Serine Protease. These properties suggest that alkaline protease from A. tamarii URM4634 is suitable for application in food industries and leather processing. Additionally, the present findings opened new vistas in the utilization of wheat bran and other effective agroindustrial wastes as substrates for SSF.

Direct Solid-State Fermentation of Soybean Processing Residues for the Production of Fungal Chitosan by Mucor rouxii

The feasibility of utilizing soybean-processing residues such as soybean meal and hulls as substrates for chitosan production by the fungus Mucor rouxii ATCC 24905 via solid-state fermentation (SSF) was investigated. The effects of the type of soybean-based substrate, length of cultivation period, substrate moisture content, substrate pH, incubation temperature and extraction conditions on chitosan yield were determined. The results showed that a maximum fungal chitosan yield of up to 3.44% by dry substrate weight (34.4 g/kg) could be achieved using a pure soybean meal substrate with an initial moisture content of 50% (w/w) and pH of 5 - 6 incubated for six days at 25&degC. A more severe heat treatment (autoclaving vs. refluxing) resulted in higher chitosan extraction yields regardless of the strength of extraction reagents. Fourier transform infrared (FTIR) analysis of the fungal chitosan revealed its degree of deacetylation (DDA) to be between 55% and 60%.

Solid-State Fermentation for the Concomitant Production of δ-Endotoxin and Endospore from <i>Bacillus thuringiensis</i>subsp. <i>kurstaki</i>

Water stress and limited aeration imparted by solid-state fermentation (SSF) were reported as crucial factors for the enhancement of endospore production by Bacillus thuringiensis (Bt);and thus, more δ-endotoxin could be produced concomitantly with reduced time. Therefore, Bt subsp. kurstaki (Btk) was employed in the present study to evaluate its efficiency for the concomitant production of endospores and δ-endotoxin in LB medium supplemented with various naturally available agricultural products, i.e., flours of soybean, Bengal gram or jack seed at various concentrations (10%, 20%, 30%, 40%, 50%, 60%, 80% or 100%, all w/v). After 12 h fermentation, the supernatant in it was centrifuged off aseptically to obtain solid substrate for subsequent SSF. Of them, soybean (30%) supplemented medium was the best for the enhanced production of endospore and δ-crystals. The maximum yield of endospores during solid-state fermentation was observed 48 h, i.e., compared to submerged fermentation in LB, it was 24 h less gestation period. In control sample, the endospores achieved the maximum length (1.10 ± 0.13 μm) and diameter (0.63 ± 0.07 μm) at 72 h;while in soybean supplemented medium, the maximum length (2.10 ± 0.16 μm) and diameter (1.63 ± 0.16 μm) were at 48 h and 72 h, respectively. Upon staining, acridine orange specifically stained the endospores;malachite green-saffranin stained both δ-crystals and endospores;and coomassie brilliant blue specifically stained δ-endotoxin. Briefly, normal gestation period or harvest time for Btk is 72 h, which could be reduced to 48 h, if SSF is employed as demonstrated in this study.

Study on Temperature Gradients and Protein Enrichment by <i>Aspergillus oryzae</i>in Solid-State Fermentation on Packed Bed Bioreactor Using Jowar (Sorghum) Straw as Substrate

The packed bed solid state bioreactor designated as PBSSB is constructed in the present study. The experiments are carried out in packed bed bioreactor with jowar straw and inoculated with Aspergillus oryzae. Temperature gradient has been measured at different axial positions. It is found that the organisms grew rapidly during the period from 20 to 30 h during which heat generation is more. These results are in agreement with other researchers. The fermented jowar straw shows threefold increase in protein content. This can be utilized as high value nutritional feed to animals.

Protein Enrichment of Potato Peels Using Saccharomyces cerevisiae via Solid-State Fermentation Process

In order to add value to potato peels and also curb their environmental pollution problems, this study investigated the protein enrichment of potato peels with Saccharomyces cerevisiae via Solid-State Fermentation (SSF). SSF is a fermentation process which involves solid matrix and is carried out in absence or near absence of free water. SSF of potato peel mashed was carried out with S. cerevisiae at 30°C, pH of 5.5, moisture adjustment between 40 and 90%, addition of ammonium sulphate and urea salts as nitrogen supplements for the microorganisms for 3 days. The results showed that the percentage crude protein content of all the fermented samples increased significantly when compared with the unfermented sample. 40% moisture content adjustment and ammonium sulphate as nitrogen source gave the best result. The crude protein increased from 12.5% to 21.86%, which is 74.88% increment for ammonium sulphate supplementation, and 12.5% to 18.42%, which is 47% increment for urea supplementation. Therefore, the fermented peels could serve as good source of cheap protein enriched feed for livestock.

Optimization of the Chitinase Production by Different Metarhizium anisopliae Strains under Solid-State Fermentation with Silkworm Chrysalis as Substrate Using CCRD

Entomopathogenic fungi, such as Metarhizium anisopliae, are able to control various insect pests. These fungi attack the integument of the host using an enzymatic complex. Among the enzymes found in this complex, chitinase is an important component. However, the relation between the chitinase production and the virulence from different M. anisopliae strains has not been analyzed. In this manuscript it is presented the chitinase production by four M. anisopliae strains with different potential of virulence in Solid-State Fermentation using silkworm chrysalis as substrate. The higher chitinase level was obtained with the strain IBCB 360 (7.14 U/g of substrate) with potential virulence of 68% on Diatrea saccharalis. The enzyme production was optimized for all strains using a factorial planning (CCRD) considering the cultivation time and medium humidity as independent variables. The maximal production of chitinase was obtained at a range from 8 to 12-days old cultures and from 45% to 62% of moisture according to the surface response plot, with high R2 value. The enzyme production by the strain IBCB 167 was increased two-folds under optimized conditions, while for the strains IBCB 360 and 425 the chitinase production was increased four-folds and nine-folds for the strain IBCB 384.

In-depth observations of fermentative hydrogen production from liquid swine manure using an anaerobic sequencing batch reactor

In this study,experiments were designed to reveal in-depth information of the effect of pH and hydraulic retention time（HRT）on biohydrogen fermentation from liquid swine manure supplemented with glucose using an Anaerobic Sequencing Batch Reactor（ASBR）System.Five values of HRT（8,12,16,20,and 24 h）were first tested and the best HRT determined was further studied at five p H levels（4.4,4.7,5.0,5.3,and 5.6）.The results showed that for HRT 24 h,there was a dividing H2 content（around 37%）related to the total biogas production rate for the ASBR System running at p H 5.0.When the H2 content went beyond 37%,an appreciable decline in biogas production rate was observed,implying that there might exist an H2 content limit in the biogas.For other HRTs（8 through 20 h）,an average H2 content of 42%could be achieved.In the second experiment（HRT 12 h）,the highest H2 content（35%）in the biogas was found to be associated with p H 5.0.The upswing of p H from 5.0 to 5.6 had a significantly more impact on biogas H2 content than the downswing of p H from5.0 to 4.3.The results also indicated good linear relationships of biogas and H2 production rates with HRT（r=0.9971 and0.9967,respectively）.Since the optimal ASBR operating conditions were different for the biogas/H2 production rates and the H2 yield,a compromised combination of the running parameters was determined to be HRT 12 h and pH 5.0 in order to achieve good biogas/H2 productions.

Suspensions Derived from Anaerobically Fermented Tilapia Offal Inhibit <i>Fusarium oxysporum</i>f. sp. <i>cubense</i>in Banana

Effective control strategies are lacking for Fusarium wilt of banana crops worldwide. Here, the inhibitory efficacy of suspensions derived from Tilapia offal by anaerobic fermentation against Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) was evaluated. Two anaerobic fermentation methods were used: 1) natural fermentation of offal (NF) and 2) fermentation of offal supplemented with 5% lime (LF). The suspensions were applied in three treatments: Plate assay, pot system, and in the field. The inhibition rate and disease index were determined. The results showed that the inhibition was significantly greater for LF than for NF on plates. In pot system and in the field, the disease index was lower for the LF group than for the NF group and was significantly lower than that of the control. Therefore, suspensions derived from anaerobically fermented offal provide a new control method for Fusarium wilt in banana.

Control of fermentation types in continuous-flow acidogenic reactors:effects of pH and redox potential

The experiments were carried out in continuous flow acidogenic reactors with molasses used as substrate to study the effects of pH and redox potential on fermentation types. The conditions for each fermentation type were investigated at different experimental stages of start up, pH regulating and redox potential regulating. The experiments confirmed that butyric acid type fermentation would occur at pH > 6, the propionic acid type fermentation at pH about 5.5 with E h> -278 mV, and the ethanol type fermentation at pH < 4.5. A higher redox potential will lead to propionic acid type fermentation because propionogens are facultative anaerobic bacteria.

Performance of Biogas Production from Anaerobic Codigestion of Dead Pigs and Pig Manure

With the development of large-scale and intensive pig breeding,the harmless disposal of dead pigs has become increasingly prominent.The method to process dead pigs by biogas engineering can realize resource utilization of dead pigs and and obtain high economic benefit. In this paper,through a batch of experiments,the feasibility of anaerobic co-digestion of dead pigs and pig manure was verified,and effects of addition proportion of dead pigs and reaction temperature on biogas production from the anaerobic co-digestion were analyzed. The results showed that when the reaction temperature was 35 ℃ and the adding proportion of the dead pig was 12%,the fermentation could get the best effect,and the maximum of methane content could reach 64. 72%,while the total biogas yield was up to 5 310 ml,which was 26% higher than single fermentation of pig manure. Meanwhile,the unit TS and VS biogas yield could rise by 30% and 22% respectively.

Optimization of Parameters for the Production of Lipase from Pseudomonas sp. BUP6 by Solid State Fermentation

Solid-state fermentation (SSF) holds tremendous potentials for the production of industrially significant enzymes. The present study describes the production of lipase by a novel rumen bacterium, Pseudomonas sp. strain BUP6 on agro-industrial residues. Pseudomonas sp. strain BUP6 showed higher lipase production when grown in Basal salt medium (BSM) supplemented with oil cakes. Initially, five different oil cakes (obtained after extracting oil from coconut, groundnut, cotton seed, gingelly or soybean) were screened to find out the most suitable substrate-cum-inducer for the production of lipase. Among them, groundnut cake supported the maximum production of lipase (107.44 U/gds). Box-Behnken Design (BBD), followed by response surface methodology (RSM) was employed to optimize the culture parameters for maximizing the production of lipase. Using the software Minitab 14, four different parameters like temperature, pH, moisture content and incubation time were selected for the statistical optimization, which resulted in 0.7 fold increase (i.e., 180.75 U/gds) in production of lipase under the optimum culture conditions (temperature 28&#176C, pH 5.9, moisture 33% and incubation 2 d). Thus, this study signifies the importance of SSF for the production of industrially-significant lipase using agro-industrial residues as solid support.

Coupling System for Food Wastes Anaerobic Digestion and Polyhydroxyalkanoates Production with Ralstonia eutropha

A new technology was developed to couple the anaerobic digestion of food wastes with production of polyhydroxyalkanoates (PHAs). Acetic, propionic, butyric and lactic acids were produced during food wastes anaerobic digestion and their concentrations reached 5.5, 1.8, 27.4 and 32.7 g/L, respectively under appropriate digestion conditions. The fermentative acids were transferred through a dialysis membrane to an air-lift reactor for PHA synthesis by Ralstonia eutropha. Dry cell concentration and PHA content reached 22.7 g/L and 72.6%, respectively. The obtained PHA was a copolymer of b-hydroxybutyrate (HB) and b-hydroxyvalerate (HV) with 2.8% (mole ratio) of HV units in polymer.