Study on the Fermentation Conditions of a Mucopolysaccharide-producing Bacterium LV-1

bacterium LV-1 which isolated from soil sample were studied bacterium was isolated by serial dilution method, the effects of carbon source, nitrogen source, the initial pH and temperature on producing polysaccharide by it were discussed to confirm the optimum fermentation conditions. [ Result] The physicochemical properties showed that the polysaccharide was water-soluble, but insoluble in organic solvents including ethanol, butanol, and chloroform. It was neutral polysaccharide with negative charge and without reducing terminal. The pH of its solution was pH =7.5. There were no protein, fructose, uronic acid, sulphate and starch-like structure included in po/ysacchadde molecules. The optimum fermentation conditions for po/ysaccharide produc- tion were 3% mannitol as carbon source, 0.25% yeast extract as nitrogen source, culture temperature 28 ~（3 and pH =7.5. [Coadu^en] The re- search could provide basis for development and utilization of LV-1 and industrialized production of mucopolysaccharide.

Study on Fermentation Conditions of Pycnoporus sanguineus

[Objective] The purpose was to study the optimum conditions of Pycnoporus sanguineus for producing ligninase. [Method] A strain of lignindegrading white-rot fungus was selected from 5 strains of collected fungi and its ligninase production and the optimum conditions for producing ligninolytic enzyme were measured. [Result] It could produce two kinds of ligninase with good thermal stability. Different temperatures, carbon sources, nitrogen sources, acidities, as well as the additions of surfactant had distinct influence on the development of lignin-degrading enzymes of the fungus. The optimum condition was drawn out：38℃, pH = 4.5, 10.0 g/L glucose, 1.0 g/L tartaric acid ammonium. [Conclusion] The aim of research was to provide a basis for lignin degradation in practical production.

Research on the Fermentation Conditions of Bacillus subtillus B-332

[Objective] The paper was to ascertain the fermentation conditions and medium formula of Bacillus subtilis B-332. [Method] Using single factor and orthogo- nal experimental designs, the fermentation medium and culture conditions for B. sub- i tills B-332 were optimized through shaking flask culture. [Result] The optimal culture and inoculation time was 18 h, and the optimized medium formula was as follows： soybean powder 0.60 g/L, sucrose 0.25 g/L, ammonium sulfate 0.07 g/L, trisodium citrate 0.03 g/L, dipotassium hydrogen phosphate 0.003 g/L, magnesium sulfate 0.005 g/L, ferrous sulfate 0.000 5%; the fermentation condition was as follows： tem- perature 30 ℃, shaking bed speed 180 r/min, shaking bottle volume 80 ml/500 ml. In the optimal fermentation conditions, the fermented spore quantity was 1.43×10^11 cfu/ml, which was 34.48 times higher than the spore quantity of 4.03×10^9 cfu/ml in initial condition. [Result] The study laid the foundation for factory production of the strain.

Optimization of Fermentation Conditions of Acid Resistant α-amylase Producing Strain

[Objective] The aim was to optimize the fermentation conditions of acid resistant α-amylase producing strain. [Method] Based on the selection of an acid resistant α-amylase producing strain,the fermentation conditions including C,N contents and initial pH of culture medium,seed age,inoculum size,rotation speed of shake flask and fermentation temperature were optimized. [Result] The optimum fermentation conditions for acid resistant α-amylase producing strain were：seed age 14 h,inoculum size 8%,initial pH 5.5,fermentation temperature 35 ℃,rotation speed 150 r/min,the volume of inoculum broth 25 ml,C content 1.0% and N content 0.5%. [Conclusion] Under the optimum fermentation conditions,α-amylase activity reached 31.4 U/ml,which was 65.3 % higher than that before optimization.

Study on Fermentation Conditions of Trichoderma aureoviride sp.for the Production of Chitinase

[Objective] The paper was to study the best fermentation conditions of Trichoderma aureoviride sp. for the production of chitinase, thus provide new enzyme source for chitinsse industry. [Method] Using orthognnal experimental design, with the variation of sugar content after enzymatic hydrolysis measured by DNS method as the indicator, the fermentation conditions were optimized. [Result] Taking colloidal chitin as the carbon source and 2% peptone as the nitrogen source with the shaking speed of 170 r/rain, the optimum fermentation conditions of T. aureoviride for the production of chitinase were as follows ： initial pH value of medium, 5.0; inoculation amount, 8% ; bottle volume, 20 ml; 6 d cultivation at 28℃. [Conclusion] The optimum conditions for the production of chitinase were confirmed, which provided basis for the utilization of T. aureoviride.

Study on Sporulation Conditions of Trichoderma reesei by Solid Fermentation

[Objective] This study aimed to investigate the sporulation conditions of Tri- choderma reesei by solid fermentation. [Method] With sporulation yield as the response value, single-factor test, Plackett-Burmam design, steepest ascent test, BoxBehnken design and response surface analysis were employed to optimize the con- ditions for sporulation of Trichoderma reesei by solid fermentation. [Result] Based on single-factor test, the most appropriate carbon source for Trichoderma reesei was straw stalk powder and wheat bran with the ratio of 3：2 and optimal amount of 15 g/L; the most appropriate inorganic nitrogen was （NH4）2O4 with the optimal amount of 3 g/L. According to Plackett-Burmam design, moisture content, initial pH and incubation temperature were identified as significant factors affecting the sporulation yield of Trichoderma reeseL The maximum sporulation yield area was approached by steepest ascent test. Based on Box-Behnken design and response surface analysis, the optimal fermentation conditions for the maximum sporulation yield were determined as： straw stalk powder of 6 g/L, wheat bran of 9 g/L, （NH4）2SO4 of 3 g/L, moisture content of 65%, incubation temperature of 29 ℃, fermentation period of 72 h and initial pH of 5.5, under these conditions, the sporulation yield reached 2×10^10 spores/g, which was improved by 1.4 times compared with that before optimization. [Conclusion] In this study, the conditions for sporulation of Trichoderma reesei by solid fermentation were optimized with low-cost straw stalk powder and wheat bran as carbon sources, which was conducive to reducing the production cost of Trichoderma reesei and increasing the sporulation yield, showing certain social and economic significance.

Optimization of Fermentation Medium and Conditions of Antibiotic Active Substances Produced by Antarctic Psychrotrophic Bacterium Rheinheimera sp. 97

[Objective] The aim was to optimize the fermentation medium and conditions of antibiotic active substances produced by Antarctic psychrotrophic bacterium Rheinheimera sp.97.[Method] Single-factor experiment and orthogonal test were adopted to optimize the fermentation medium of antibiotic active substances produced by Antarctic psychrotrophic bacterium R.sp.97,while the fermentation conditions were optimized by single-factor experiment.[Result] The optimum fermentation medium for the antibiotic active substances production was as follows：tryptone 3.0 g/L,ammonium sulfate 1.0 g/L,starch 2.0 g/L,NaCl 15.0 g/L.The optimized fermentation conditions were as follows：the starting pH of medium was 8.0,fermentation temperature was 10 ℃,liquid volume in Erlenmeyer flask was 30 %（V/V）and inoculation amount was 1%（V/V）.Under the optimized fermentation medium and conditions,the antibacterial activity of R.sp.97 was increased by 18.1%.[Conclusion] This study had provided basis for the antibiotic active substances produced by Antarctic psychrotrophic bacterium R.sp.97.

Optimization of the Fermentation Conditions of Sweet Oats by Response Surface Methodology

Objective] The purpose of this study was to evaluate the effects of five variables （steaming time, moisture content before inoculation, inoculation amount, fer-mentation temperature, fermentation time） on the sweetness of sweet oats, obtained the best fermentation conditions. [Method] Plackett-Burman （PB） was to evaluate the effects of five variables, and selected significant factors. The steepest ascent was used to approach the optimal response surface experimental area. The optimal fer-mentation condition was obtained by central composite design and response surface analysis. [Results] It was indicated that moisture content, inoculation amount and fer-mentation temperature had significant influences on the content of the reducing sugars. The optimal conditions of moisture content, inoculation amount and temper-ature were 45.26%, 0.014%（g/g） and 28 ℃, respectively. The predicted value of the reducing sugar content was 13.16 mg/g. [Conclusion] Under the optimal conditions, the content of the reducing sugars in the sweet oats could be up to 12.91 mg/g, which was on the whole consistent with the predictive maximum value.

Optimization of Fermentation Conditions for Xylanase Production by Aspergillus niger NS-1

In this study, a xylanase-produeing Aspergillus niger strain, NS-1, was screened and isolated from agricultural and forestry wastes. Based on single-fac- tor experiments, the effects of different carbon sources, composite carbon sources, nitrogen sources, calcium carbonate concentrations, initial pH and surfactants on xylanase production by A. niger NS-1 were investigated. The results indicated that the most appropriate carbon source was corncobs ; the best composite carbon source was corncobs ＋ xylan, which was conducive to xylanase secretion; the most suitable nitrogen source was ammonium sulfate. Xylanase activity reached the highest in the medium added with 1.5% calcium carbonate and SDS as a surfactant with an initial pH of 5.0. This study provided the basis for the production of high-activity xylanase.

Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain ,Aspergillusfoetidus ZU-GI in solid-state fermentation （SSF）. Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60% initial moisture of medium, 28 ℃ incubation temperature, 18^h cultivation period of seed, 10% inoculum volume, 5.0-6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2037.51 U/g dry matter near the 144th hour of fermentation.

Herbicidal activity of Aureobasidium pullulans PA-2 on weeds and optimization of its solid-state fermentation conditions

Fungal strain PA-2 was isolated from infected poplar leaves from the Ping’an District, Haidong City of Qinghai Province, China. Based on the culture characteristics and the internal transcribed spacer(ITS) sequence of its 16S r DNA, the strain was identified as Aureobasidium pullulans. The culture and metabolites of strain PA-2 showed high herbicidal potential to five tested weeds Galium aparine var. tenerum, Chenopodium album, Malva crispa, Polygonum lapathifolium and Avena fatua. For the in vitro test, 5 days after the detached leaves were inoculated with PA-2 culture, all leaves infected by the hyphae and became black and rotten. For the in vivo test, a metabolite filtrate of PA-2 culture was sprayed over the living weed plants, and five days after inoculation, the weed plants became withered and necrotic. Seven days after inoculation, the fresh weight reductions of G. aparine var. tenerum, C. album, M. crispa, P. lapathifolium and A. fatua were 87.25, 78.46, 82.25, 62.11, and 80.27%, respectively. Galium aparine var. tenerum and M. crispa exhibited significant reductions in fresh weight. The bio-safety test on the five crops showed no significant plant height reductions, which was also observed for wheat(Triticum aestivum), faba bean(Vicia faba), and barley(Hordeum vulgare). By contrast, oilseed rape(Brassica napus) and pea(Pisum sativum) exhibited light spots but no significant reductions in plant height. These results indicated that A. pullulans could be a potential microbial herbicide for the control of the target weeds in crops. Optimization of the carbon and nitrogen sources for cultural media and substances for solid-state fermentation indicated that PA-2 had better colony growth and spore production with the optimal carbon source of glucose(C4), nitrogen source of soybean flour(N2), and the optimal substance was wheat bran. The results in this study provide useful information for the development ofA. pulluans PA-2 as an herbicide for bio-control of the weed.

Nutrient Requirements and Fermentation Conditions for Mycelia and Crude Exo-Polysaccharides Production by <i>Lentinus squarrosulus</i>

Lentinus squarrosulus Mont. is an emerging tropical white rot basidiomycete, with nutritional and medicinal benefits. Low levels of commercial cultivation of the mushrooms limit their availability for use as food and medicine. Mycelia from submerged fermentation are a suitable alternative to the mushroom from L. squarrosulus. Three strains, 340, 339 and 218, were studied to determine optimum growth conditions for mycelia mass and crude exo-polysaccharides (CEPS) production. The experiments were conducted in a completely randomized design (CRD) with a factorial structure. Nutrients involving 8 carbon and 8 nitrogen sources were screened, and concentrations of the best sources were optimized. Optimized nutrients, interaction between strains and other parameters such as agitation and medium volume were investigated to obtain optimum fermentation conditions for biomass and CEPS production. Biomass yield varied among strains depending on carbon or nitrogen nutrient sources. Starch and yeast extract at 30 and 25 g/L were identified as the most important nutrients in mycelia and CEPS production. Nutrient optimization resulted in a 3-fold increase in mycelia mass: 12.8, 10.0 and 15.3 g/L in strains 340, 339 and 218 respectively. There was a significant interaction between strain, agitation, and volume (p p = 0.02). Static conditions favored more polysaccharide production. Optimized fermentation conditions resulted in very high increase in biomass: 238.1, 266.9 and 185.0 g/L in strains 340, 339 and 218 respectively. Results obtained could be useful in modeling fermentation systems for large-scale production of mycelia mass, CEPS and other bio-products from L. squarrosulus.

Optimization of Fermentation Conditions of Lepista sordida Mycelium

[ Objective] This study aimed to optimize the fermentation condition of Lepista sordida mycelium. [ Method ] The effects of carbon sources, nitrogen sources ,pH and incubation time on the dry weight of mycelium in L. sordida fermentation were investigated. [ Result] The descending order of the influences on L. sordida mycelium growth was ： carbon sources 〉 incubation time 〉 pH 〉 nitrogen sources. Based on orthogonal experiment, the optimal fermentation condition of L. sordida mycelium was determined as ： fermentation medium of 3% corn powder and 0.2% yeast extract, pH of 6.0, and incubation time of 8 d. [ Conclusion ] This study provides theoretical reference and experimental basis for the development and utilization of L. sordida biological resources.

Isolation of Cordyceps ophioglossoides L2 from Fruit Body and Optimization of Fermentation Conditions for Its Mycelial Growth

A strain isolated from the fruiting body of a fungus parasitized on Elaphomyces was identified as Cordyceps ophioglossoides based on the morphological characteristics and the analysis of ITS-5.8s rDNA sequence. The optimal medium, composition （g·L^-1）, containing sucrose 66.0, yeast powder 10.0, silkworm chrysalises digest 30.0, MgSO4· 7H2O 0.4, and KH2PO4 0.4, Was found using fractional factorial design ancl a central composite design, and the optimization of cultural conditions obtained a result of seed age 6 days, inoculum size 6% （by volume）, initial pH 5.6, temperature 24℃, shaking speed 160 ·＇min^-1 by one-factor-at-a-time method. The maximum biomass reached about 20.2 g·L^-1 after 90 hours culture under the optimal conditions. Elementary nharmaeclogical actlwtties showed that mycelia of C. ophioglossoides L2 from submerged culture promoted Uterus growth in estrogen- depleted mice. In the 15-litre scale-up fermentation, the mycelial biomass was around 19.1 g·L^-1, indicating a promising prospect for this biotechnoloagy and the potency to develoo its medical value.

Optimization of Solid State Fermentation Conditions Using a Mixture of Bean Curd Residue and Marc with Bacillus natto

[Objective] The aim was to optimize the appropriate solid state fermentation(SSF)conditions.[Method] The optimization of solid state fermentation using a mixture substrate of bean curd residue and the marc with Bacillus natto was developed.[Result] The best fermentation condition optimized by the test of single factor and the orthogonal design respectively was mixing ratio of bean curd residue to marc 2∶1,substrate pH value 6,fermentation temperature 39 ℃,inoculum volume 10% and fermentation time 48 h.Under this optimized fermentation condition,the content of crude fiber in the substrate decreased from 107.8 mg/g before SSF to 56.2 mg/g after SSF,and the degeneration rate of crude fiber was 47.87%.[Conclusion] The bean curd residue in its palatability was enormously improved by SSF with Bacillus natto strain,which could be expected to be widely used as raw material of health foodstuff.

DES Mutagenesis of Antagonistic Streptomyces of Botryosphaeria dothidea and Optimization of Its Fermentation Conditions

[ Objective] The paper wag to improve the antimicrobial effect of antagonistic Streptomyces on Botryosphaeria dothidea. [ Method ] Different concentrations of DES were adopted for mutagenic treatment of antagonistic Streptomyces F-58. After fermentation, the mutant strain numbered F-58-06 with genetic stability and good antimicrobial effect was selected among 200 mutant strains; the biological activity of the strain was determined with B. dothidea as the indicator fungus, and the best concentration of DES wag 2%. The fermentation conditions of antagonistic substances of Streptomyces strain F-58-06 produced in shake flasks were opti- mized using the method of single factor and orthogonal experimental design. [ Result] The optimal fermentation conditions were carbon source corn flour, nitrogen source KNO3, initial pH 7, loading volume 30 mL in 250 mL shake flask, inoculation volume 3%, incubation time 132 h. Based on orthogonal experiments, the best cultured medium ratio was corn flour 4%, KNO3 0.5%, CaCO3 0.2%, NaCI 0.35%, MgSO4 · 7H2O 0. 15%. [ Conclusion ] After mutation and optimization of culture medium, the antimicrobial activity of Streptomyces F-58 was greatly improved, and the average inhibition zone diameter was 1.35 times of that of the original strain.

Studies on Fermentation Conditions of Antimicrobial Substances Produced by Brevibacillus laterosporus BL-21

[Objective] Brevibacil us laterosporus BL-21 was inhibitory to Gibberel a sanbinetti, Fusarium graminearum Magnaporthe grisea and many other phy-topathogenic fungi. Antimicrobial substances can be increased through optimizing the medium of Brevibacil us laterosporus. [Method] A total of 10 factors were tested for their function of improving antibiotics yield of BL-21 by single-factor test. And the indicative bacterium was screened. [Result] The best indicative bacterium was Al-ternaria solani, the optimum process parameters in shaking flask on fermentation consisted of initial pH of 8.0, culture at 30 ℃, loadage 90 ml/250 ml, rotation 180 r/min, and fermentation time of 48 h. The results also indicated that glucose was the best carbon source for the antibiotics productivity of BL-21, and beef extract was the best nitrogen source. In addition, univalent ion Cl＋ and divalent ion Mn2＋ stimu-lated BL-21 antibacterial active component biosynthesis. [Conclusion] Optimized medi-um is more conducive to produce antimicrobial substances.

Optimization of fermentation conditions for 1,3-propanediol production by marine Klebsiella pneumonia HSL4 using response surface methodology

The industrially important organic compound 1,3-propanediol （1,3-PDO） is mainly used as a building block for the production of various polymers. In the present study, response surface methodology protocol was followed to determine and optimize fermentation conditions for the maximum production of 1,3-PDO using marine-derived Klebsiella pneumoniae HSL4. Four nutritional supplements together with three independent culture conditions were optimized as follows： 29.3 g/L glycerol, 8.0 g/L K2HPO4, 7.6 g/L （NH4）2SO4, 3.0 g/L KH2PO4, pH 7.1, cultivation at 35℃ for 12 h. Under the optimal conditions, a maximum 1,3-PDO concentration of 14.5 g/L, a productivity of 1.21 g/（L＇h） and a conversion of glycerol of 0.49 g/g were obtained. In comparison with the control conditions, fermentation under the optimized conditions achieved an increase of 38.8% in 1,3-PDO concentration, 39.0% in productivity and 25.7% in glycerol conversion in flask. This enhancement trend was further confirmed when the fermentation was conducted in a 5-L fermentor. The optimized fermentation conditions could be an important basis for developing low- cost, large-scale methods for industrial production of 1,3-PDO in the future.

Insights into microbiota community dynamics and flavor development mechanism during golden pomfret(Trachinotus ovatus)fermentation based on single-molecule real-time sequencing and molecular networking analysis

Popular fermented golden pomfret(Trachinotus ovatus)is prepared via spontaneous fermentation;however,the mechanisms underlying the regulation of its flavor development remain unclear.This study shows the roles of the complex microbiota and the dynamic changes in microbial community and flavor compounds during fish fermentation.Single-molecule real-time sequencing and molecular networking analysis revealed the correlations among different microbial genera and the relationships between microbial taxa and volatile compounds.Mechanisms underlying flavor development were also elucidated via KEGG based functional annotations.Clostridium,Shewanella,and Staphylococcus were the dominant microbial genera.Forty-nine volatile compounds were detected in the fermented fish samples,with thirteen identified as characteristic volatile compounds(ROAV>1).Volatile profiles resulted from the interactions among the microorganisms and derived enzymes,with the main metabolic pathways being amino acid biosynthesis/metabolism,carbon metabolism,and glycolysis/gluconeogenesis.This study demonstrated the approaches for distinguishing key microbiota associated with volatile compounds and monitoring the industrial production of high-quality fermented fish products.

Effect of Corn Stalks Treated with Carbonylamines in Different Concentrate and Forage Rations and Nitrogen Conditions on Rumen Fermentation in Vitro

[ Objective] To investigate the effects of concentrate/roughage ratios and nitrogen levels on in-vitro rumen fermentation of urea-treating corn stalk. [ Method] The concentrate/roughage ratios were 7： 3, 5：5 and 3： 7. The crude protein levels were 11% and 14%. The in-vitro culture time was 2, 4, 6 and 48 h. [ Result] The pH value of broth decreased significantly with the decline in the proportion of roughage （ P 〈 0.01 ） and with the increase in the crude protein levels （P 〈 0.05）. The ammonium nitrogen concentration and acetic acid/propionic acid ratio of the fermen- ted products decreased with the increase in the crude protein levels and proportion of concentrate in diet. The diet with concentrate/roughage ratio of 5：5 and crude protein level of 14% had significantly higher digestibility of dry matter and organic matter than other diets （ P 〈 0.05）. The digesti- bility of organic matter increased gradually with the increasing proportion of concentrate and crude protein level. With the increase in the proportion of concentrate, the microbial protein levels increased remarkably, while the acetic acid/propionic acid ratio declined. [ Conclusion] The concentrate/ roughage ratios and nitrogen levels affect rumen fermentation and microbial growth during in-vitro culture. However, the best supplementary feeding results of urea-treatinq corn stalks can be obtained when the concentrate/rouahaae ratio is below 5：5 and the crude protein level is 14%.