Growth ofCandida tropicalis 2838 cells on straw hydrolyzates

Improved strain of Candida tropicalis 2838 grows on nonseparated straw hydrolyzates with no addition of vitamins and trace elements at a specific growth rate mu = 0.34 and 44% yield coefficient (referred to reducing substances). The reducing substances in hydrolyzates contain predominantly monosaccharides (xylose, glucose, arabinose, mannose). Cells grown in this way are rich in proteins (62%) and essential amino acids (lysine, phenylalanine, leucine, threonine and valine). The product obtained under industrial conditions by fermentation of the nonseparated hydrolyzates contains 8--9% of proteins and it is a suitable supplement of fodder mixtures for monogastric domestic animals. Nutrition tests on rats and pigs indicated that this product can substitute the hay-flour, and, partially, blood-flour barley, and that the strain used is nonpathogenic.     

The use of cellulases from a beta-glucosidase-hyperproducing mutant of Trichoderma reesei in simultaneous saccharification and fermentation of wheat straw

Conidia of the cellulolytic strain Trichoderma reesei F522 were mutagenized with UV irradiation and N-methyl|-N'-nitro-N-nitrosoguanidine (NTG). A visual agar plate detection system was developed, using esculin and ferric ions, to identify mutants of T. reesei with increased beta-glucosidase activity. Selected mutants were tested for production of extracellular cellulases in shake flasks on autohydrolyzed wheat straw as carbon source. The most active mutant V-7 showed about 6-times higher activity of beta-glucosidase than the parent strain F-522, whereas the filter paper degrading and endo-1,4-beta-D-glucanase activities increased by 45% and by almost 31%, respectively. Cellulase preparations obtained from the parent and mutant strains were then used along with Kluyveromyces fragilis cells for ethanol production from ethanol-alkali pulped straw in the simultaneous saccharification and fermentation (SSF) process. From 10% (w/v) of straw pulp (dry matter), 2.5% (w/v) ethanol was obtained at 43 degrees C after 48 h using cellulase derived from the parent strain of T. reesei. When the beta-glucosidase-hyperproducing mutant V-7 was employed, the ethanol yield in the SSF process increased to 3.4% (w/v), the reaction time was shortened to 24 h and no cellobiose was detected in straw hydrolyzates.     

Optimization of the pretreatment of rice straw hemicellulosic hydrolyzates for microbial production of xylitol

Hemicellulosic hydrolyzate obtained from rice straw was evaluated to determine if it was a suitable fementation medium for the production of xylitol byCandida mogii ATCC 18364. To obtain xylose selectively from rice straw, it is important to establish rapid hydrolysis conditions that yield xylose-rich substrates. The results of hydrolysis experiments indicated that the optimal reaction conditions for the recovery of xylose from rice straw hemicellulose were obtained using a sulfuric acid concentration of 1.5%, a reaction temperature of 130°C, a reaction time of 20 min and a solid to liquid ratio of 1∶10. Because the fermentation of concentrated acid hydrolyzates can be inhibited by compounds present in the raw material or produced during the hydrolysis process, various methods were tested to determine if they could detoxify the hydrolyzates and thus improve xylitol production. The greatest xylitol yield (0.53 g/g) and volumetric productivity (0.38 g/L·h) were obtained when an overlimed hydrolyzate was treated with activated charcoal.     

Dilute-acid hydrolysis for fermentation of the Bolivian straw material Paja Brava

Hydrolysis of the straw material Paja Brava, a sturdy grass characteristic for the high plains of Bolivia, was studied in order to find suitable conditions for hydrolysis of the hemicellulose and cellulose parts. Dried Paja Brava material was pre-steamed, impregnated with dilute sulfuric acid (0.5% or 1.0% by wt), and subsequently hydrolyzed in a reactor at temperatures between 170 and 230 degrees C for a reaction time between 3 and 10 min. The highest yield of xylose (indicating efficient hydrolysis of hemicellulose) were found at a temperature of 190 degrees C, and a reaction time of 5-10 min, whereas considerably higher temperatures (230 degrees C) were needed for hydrolysis of cellulose. Fermentability of hemicellulose hydrolyzates was tested using the xylose-fermenting yeast species Pichia stipitis, Candida shehatae and Pachysolen tannophilus. The fermentability of hydrolyzates decreased strongly for hydrolyzates produced at temperatures higher than 200 degrees C.     

Utilization of agro-industrial waste for xylanase production by Aspergillus foetidus MTCC 4898 under solid state fermentation and its application in saccharification

Xylanase production by Aspergillus foetidus MTCC 4898 was carried out under solid state fermentation using wheat bran and anaerobically treated distillery spent wash. Response surface methodology involving Box–Behnken design was employed for optimizing xylanase production. The interactions among various fermentation parameters viz. moisture to substrate ratio, inoculum size, initial pH, effluent concentration and incubation time were investigated and modeled. The predicted xylanase activity under optimized parameters was 8200–8400 U/g and validated xylanase activity was 8450 U/g with very poor cellulase activity. Crude xylanase was used for enzymatic saccharification of agroresidues like wheat straw, rice straw and corncobs. Dilute NaOH and ammonia pretreatments were found to be beneficial for the efficient enzymatic hydrolysis of all the three substrates. Dilute NaOH pretreated wheat straw, rice straw and corncobs yielded 4, 4.2, 4.6 g/l reducing sugars, respectively whereas ammonia treated wheat straw, rice straw and corncobs yielded 4.9, 4.7, 4.6 g/l reducing sugars, respectively. The hydrolyzates were analysed by HPTLC. Xylose was found to be the major end product with traces of glucose in the enzymatic hydrolyzates of all the substrates.     

一株促生秸秆降解菌的分离与鉴定

针对秸秆处理不当影响全世界环境污染的问题，筛选多功能秸秆降解菌，旨在得到高效降解秸秆且具有促生作用的微生物菌种。结合纤维素钠-刚果红(CMC-Na)平板筛选，通过16S rRNA基因分析，进行菌株鉴定，得到一株具有纤维素降解效果的菌株XJ-132,经16S rRNA基因鉴定为枯草芽胞杆菌(Bacillus subtilis)。与单独施用秸秆处理相比，加入菌株XJ-132 60 d后，秸秆降解率提高21.0%,且对水稻生长促进作用显著，地上、下部鲜重分别增加17.8%和9.6%。水稻种子喷施菌株XJ-132发酵液，低浓度发酵液对种子萌发具有一定促进作用。结果表明，菌株XJ-132可能通过产吲哚乙酸(IAA)、产铁载体、产氨等多种有益物质，降解秸秆的同时促进水稻生长。筛选具有促生作用的秸秆降解菌能够更好地加速秸秆降解，具有广泛的开发利用前景。     

Description of <Emphasis Type="Italic">Streptomyces neopeptinius</Emphasis> sp. nov., an actinobacterium with broad spectrum antifungal activities

A streptomycete strain producing broad-spectrum antifungal substances was taxonomically characterized. The strain, designated KNF 2047(T) (= SH-09(T) = KCTC 10586BP(T)), was found to form extensively branching aerial and substrate mycelia, and produce spiny-ornamented spores with loose spiral chains. The whole cell hydrolyzates contained major amount of LL-diaminopimelic acid. The major fatty acids of the phospholipids were saturated and branched fatty acids containing 14~17 carbons, and the major isoprenoid quinones were hexa-and octa-hydrogenated menaquinones with 9 isoprene units. The phylogenetic analysis using the 16S rRNA gene indicated that the strain belongs to the genus Streptomyces but forms an independent phyletic line. These results clearly demonstrate that strain KNF2047(T) forms a new center of taxonomic variation within Streptomyces, for which the name Streptomyces neopeptinius sp. nov. is proposed.     

Bioconversion of wheat straw cellulose/hemicellulose to ethanol by Saccharomyces uvarum and Pachysolen tannophilus

The information presented in this publication represents current research findings on the production of glucose and xylose from straw and subsequent direct fermentation of both sugars to ethanol. Agricultural straw was subjected to thermal or alkali pulping prior to enzymatic saccharification. When wheat straw (WS) was treated at 170 degrees C for 30-60 min at a water-to-solids ratio of 7:1, the yield of cellulosic pulp was 70-82%. A sodium hydroxide extration yielded a 60% cellulosic pulp and a hemicellulosic fraction available for fermentation to ethanol. The cellulosic pulps were subjected to cellulase hydrolysis at 55 degrees C for production of sugars to support a 6-C fermentation. Hemicellulose was recovered from the liquor filtrates by acid/alcohol precipitation followed by acid hydrolysis to xylose for fermentation. Subsequent experiments have involved the fermentation of cellulosic and hemicelluosic hydrolysates to ethanol. Apparently these fermentations were inhibited by substances introduced by thermal and alkali treatment of the straws, because ethanol efficiencies of only 40-60% were achieved. Xylose from hydrolysis of wheat straw pentosans supported an ethanol fermentation by Pachysolen tannophilus strain NRRL 2460. This unusual yeast is capable of producing ethanol from both glucose and xylose. Ethanol yields were not maximal due to deleterious substances in the WS hydrolysates.     

Studies on the Degradation Mechanisms of Proteins and their Derivatives in the Foods

The volatile sulfur components produced by boiling soybean meal hydrolyzates (AMINOSAN-EKI) have been identified as dimethyl sulfide and hydrogen sulfide. No mercaptan or disulfides were detected.

The main precursor of dimethyl sulfide is supposed to be methionine methylsulfonium compound derived from methionine and pectin substances (–COOCH₃) during the hydrolysis of soybean meal by hydrochloric acid.     

Potential of biotechnological conversion of lignocellulose hydrolyzates by Pseudomonas putida KT2440 as a model organism for a bio‐based economy

Lignocellulose‐derived hydrolyzates typically display a high degree of variation depending on applied biomass source material as well as process conditions. Consequently, this typically results in variable composition such as different sugar concentrations as well as degree and the presence of inhibitors formed during hydrolysis. These key obstacles commonly limit its efficient use as a carbon source for biotechnological conversion. The gram‐negative soil bacterium Pseudomonas putida KT2440 is a promising candidate for a future lignocellulose‐based biotechnology process due to its robustness and versatile metabolism. Recently, P. putida KT2440_xylAB which was able to metabolize the hemicellulose (HC) sugars, xylose and arabinose, was developed and characterized. Building on this, the intent of the study was to evaluate different lignocellulose hydrolyzates as platform substrates for P. putida KT2440 as a model organism for a bio‐based economy. Firstly, hydrolyzates of different origins were evaluated as potential carbon sources by cultivation experiments and determination of cell growth and sugar consumption. Secondly, the content of major toxic substances in cellulose and HC hydrolyzates was determined and their inhibitory effect on bacterial growth was characterized. Thirdly, fed‐batch bioreactor cultivations with hydrolyzate as the carbon source were characterized and a diauxic‐like growth behavior with regard to different sugars was revealed. In this context, a feeding strategy to overcome the diauxic‐like growth behavior preventing accumulation of sugars is proposed and presented. Results obtained in this study represent a first step and proof‐of‐concept toward establishing lignocellulose hydrolyzates as platform substrates for a bio‐based economy.     

Improving enzymatic hydrolysis of wheat straw using ionic liquid 1-ethyl-3-methyl imidazolium diethyl phosphate pretreatment

This study aims to establish a cellulose pretreatment process using ionic liquids (ILs) for efficient enzymatic hydrolysis. The IL 1-ethyl-3-methyl imidazolium diethyl phosphate ([EMIM]DEP) was selected in view of its low viscous and the potential of accelerating enzymatic hydrolysis, and it could be recyclable. The yield of reducing sugars from wheat straw pretreated with this IL at 130 °C for 30 min reached 54.8% after being enzymatically hydrolyzed for 12 h. Wheat straw regenerated were hydrolyzed more easily than that treated with water. The fermentability of the hydrolyzates, obtained after enzymatic saccharification of the regenerated wheat straw, was evaluated using Saccharomyces cerevisiae. This microbe could ferment glucose efficiently, and the ethanol production was 0.43 g/g glucose within 26 h. In conclusion, the IL [EMIM]DEP shows promise as pretreatment solvent for wheat straw, although its cost should be reduced and in-depth exploration of this subject is needed.     

秸秆降解放线菌GC的筛选及其应用基础研究

运用循环流化技术,从土壤样品中筛选出1株具有单独降解秸秆能力的菌株GC,考察了该菌的生长特性及产纤维素酶和木质素酶能力,验证了该菌对小麦秸秆的处理效果。结果表明,该菌为放线菌的左式链霉菌(Streptomyces drozdowiczii);可在LB等基础培养基中快速繁殖;纤维素内切酶活和滤纸酶活分别可达67.57 U/mL和19.69 U/mL,并且具备木质素降解能力;该菌单独处理小麦秸秆20 d的秸秆失重率为11.52%;处理产物含多种石油烃、有机醇和植物甾醇等,表明该菌在秸秆等农业面源污染物的资源化利用方面具有良好的开发应用前景。     

Effect of bio-treatment on the lipophilic and hydrophilic extractives of wheat straw

Wheat straw, an important papermaking raw material in China, was treated with a white-rot fungus of Phanerochaete chrysosporium ME446, and the lipophilic and hydrophilic extractives from the control and bio-treated samples were analyzed by GC and GC–MS. Bio-treatment of wheat straw could alter the chemical composition of both the lipophylic and hydrophilic extractives. Sugars and phenolic substances such as coniferyl alcohol, 4-hydroxycinnamic acid, 1-guaiacylglycerol and ferulic acid were substantially degraded or consumed by the fungus. More lipophilic substances such as wax, glycerides and steryl esters were degraded into the corresponding components, resulting in much higher concentrations of fatty acids and sterols in the bio-treated samples. Obviously, the bio-treatment of wheat straw was of benefit to pitch control in pulping and papermaking processes, in the view of degradation of the more lipophilic substances. In addition, the bio-treatment could increase the lignin concentration in hot-water extractives of wheat straw.     

粗糙脉孢菌中甾醇还原酶基因erg24对麦角甾醇合成的影响

粗糙脉孢菌为子囊菌中的高效纤维素降解菌,可以直接以纤维素为营养源进行生长。本研究以粗糙脉孢菌为实验对象,利用基因工程技术构建甾醇还原酶基因erg24的高表达菌株,分别以蔗糖、麦麸、玉米秸秆、小麦秸秆、杨树木屑、水稻秸秆6种物质的粉末为碳源培养野生型粗糙脉孢菌和erg24高表达菌株,利用半定量RT-PCR测定在不同培养条件下erg2、erg24和erg6 3个麦角甾醇合成相关基因的表达水平,采用HPLC方法测定不同培养条件下麦角甾醇的积累量。研究结果表明,分别以玉米秸秆、杨树木屑、水稻秸秆这3种粉末为碳源时,培养物中的erg2、erg24和erg6 3个基因表达量较高。在不同培养条件下erg24高表达菌株合成麦角甾醇量显著高于野生型粗糙脉孢菌的合成量,且以杨树木屑粉末为碳源培养时,所获得的麦角甾醇产量最高,为30.53μg/mg。结果表明erg24基因是粗糙脉孢菌合成麦角甾醇的关键基因之一,利用玉米秸秆、小麦秸秆、杨树木屑或水稻秸秆粉末为碳源培养粗糙脉孢菌时,可获得较高产量的麦角甾醇。研究结果为以农业废弃物为营养源,利用真菌生产麦角甾醇奠定了基础。     

Isolation and characterization of rice straw degrading <Emphasis Type="Italic">Streptomyces griseorubens</Emphasis> C-5

To reutilize rice straw generated during the agricultural production process, the actinomycete strain C-5 was isolated from soil that was under the stook for several years in the Heilongjiang province of China by using multiple selective culture media. Strain C-5 was identified as Streptomyces griseorubens by China General Microbiological Culture Collection Center (CGMCC) through morphological and physiological characterization combined with the result of 16S rRNA gene sequence and data analysis. This strain has simultaneous cellulase, laccase, peroxidase, xylanase and pectinase activity. The various chemical composition of rice straw were determined during fermentation process. Simultaneously the biodegradation process of rice straw stem was observed by scanning electron microscope (SEM). It is predicted that strain C-5 could decompose rice straw effectively and had promising prospects of being applied in improving the resource utilization of rice straw.     

Degradation and utilization of cellulose and straw by three different anaerobic fungi from the ovine rumen

Three different ruminal fungi, a Neocallimastix sp. (strain LM-1), a Piromonas sp. (strain SM-1), and a Sphaeromonas sp. (strain NM-1), were grown anaerobically in liquid media which contained a suspension of either 1% (wt/vol) purified cellulose or finely milled wheat straw as the source of fermentable carbon. Fungal biomass was estimated by using cell wall chitin or cellular protein in cellulose cultures and chitin in straw cultures. Both strains LM-1 and SM-1 degraded cellulose with a concomitant increase in fungal biomass. Maximum growth of both fungi occurred after incubation for 4 days, and the final yield of protein was the same for both fungi. Cellulose degradation continued after growth ceased. Strain NM-1 failed to grow in the cellulose medium. All three anaerobic fungi grew in the straw-containing medium, and loss of dry weight from the cultures indicated degradation of straw to various degrees (LM-1 greater than SM-1 greater than NM-1). The total fiber component and the cellulose component of the straw were degraded in similar proportions, but the lignin component remained undegraded by any of the fungi. Maximum growth yield on straw occurred after 4 days for strain LM-1 and after 5 days for strains SM-1 and NM-1. The calculated yield of cellular protein for strain LM-1 was twice that of both strains SM-1 and NM-1. The cellular protein yield of strain SM-1 was the same in both cellulose and straw cultures. In contrast to cellulose, straw degradation ceased after the end of the growth phase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Degradation and utilization of cellulose and straw by three different anaerobic fungi from the ovine rumen.

Three different ruminal fungi, a Neocallimastix sp. (strain LM-1), a Piromonas sp. (strain SM-1), and a Sphaeromonas sp. (strain NM-1), were grown anaerobically in liquid media which contained a suspension of either 1% (wt/vol) purified cellulose or finely milled wheat straw as the source of fermentable carbon. Fungal biomass was estimated by using cell wall chitin or cellular protein in cellulose cultures and chitin in straw cultures. Both strains LM-1 and SM-1 degraded cellulose with a concomitant increase in fungal biomass. Maximum growth of both fungi occurred after incubation for 4 days, and the final yield of protein was the same for both fungi. Cellulose degradation continued after growth ceased. Strain NM-1 failed to grow in the cellulose medium. All three anaerobic fungi grew in the straw-containing medium, and loss of dry weight from the cultures indicated degradation of straw to various degrees (LM-1 greater than SM-1 greater than NM-1). The total fiber component and the cellulose component of the straw were degraded in similar proportions, but the lignin component remained undegraded by any of the fungi. Maximum growth yield on straw occurred after 4 days for strain LM-1 and after 5 days for strains SM-1 and NM-1. The calculated yield of cellular protein for strain LM-1 was twice that of both strains SM-1 and NM-1. The cellular protein yield of strain SM-1 was the same in both cellulose and straw cultures. In contrast to cellulose, straw degradation ceased after the end of the growth phase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interaction of apolipoprotein B-containing lipoprotein secreted by Hep G2 cells with receptors for low-density lipoprotein.

Radioligand and immunoenzymatic techniques were used to characterize the receptor binding properties of apolipoprotein B-containing lipoprotein produced by HepG2 cell line (H-LpB). It was found that compared to plasma low-density lipoprotein (LDL), the interaction of H-LpB nonseparated from conditioned medium with normal fibroblasts was 6-8-times lower and only slightly exceeded the nonspecific binding of LDL modified by malondialdehyde, while the uptake of the indicated lipoproteins by LDL receptor-negative strain of fibroblasts were identical. The uptake of H-LpB by normal fibroblasts increased 1.5-2-times after isolation from the culture medium by immunoaffinity chromatography. The effect of isolation could be explained by the finding that apolipoprotein E-containing lipoprotein secreted by HepG2 cells effectively competed for the binding with LDL-receptors. The obtained results suggest that H-LpB produced by HepG2 cells is poorly recognized by the LDL-receptors.     

The role of water-soluble components in phytotoxicity from decomposing straw

Summary When seedling development is slowed by the presence of straw in wet seed-beds both microbial products and compounds of plant origin contribute to phytotoxicity. Hot (100°C) water-soluble extracts from fresh straw contained phytotoxic substances but these accounted for less of the phytotoxicity than the microbial products, primarily acetic acid, from anaerobic fermentation of the insoluble straw polysaccharides (cellulose and hemicelluloses). The water-soluble components however also included mineral salts required in the decomposition of these polysaccharides.     

Interactions between anaerobic cellulolytic bacteria and fungi in the presence of Methanobrevibacter smithii

A mixed inoculum of cellulolytic rumen bacteria depressed straw degradation by a mixed culture of cellulolytic fungi grown in the presence of Methanobrevibacter smithii. The inhibitory effect appeared to be caused by Ruminococcus albus strain JI and R. flavefaciens strain 007. Ruminococcus albus strain J1 also depressed straw degradation by the fungi, but R. albus strain SY3 and three strains of Bacteroides (Fibrobacter) succinogenes tested showed little or no inhibitory activity. It seems that some ruminococci show competitive or antagonistic activity towards certain rumen fungi.