1.Immobilization of Lactobacillus bulgaricus with gellan gum and its application in continuous fermentation of D-lactic acid from corn straw.
Yongxin GUO ; Gang WANG ; Kexin LI ; Jiaqi HAN ; Huan CHEN ; Sitong ZHANG ; Yanli LI ; Guang CHEN
Chinese Journal of Biotechnology 2023;39(3):1083-1095
Biorefinery of chemicals from straw is an effective approach to alleviate the environmental pollution caused by straw burning. In this paper, we prepared gellan gum immobilized Lactobacillus bulgaricus T15 gel beads (LA-GAGR-T15 gel beads), characterized their properties, and established a continuous cell recycle fermentation process for D-lactate (D-LA) production using the LA-GAGR-T15 gel beads. The fracture stress of LA-GAGR-T15 gel beads was (91.68±0.11) kPa, which was 125.12% higher than that of the calcium alginate immobilized T15 gel beads (calcium alginate-T15 gel beads). This indicated that the strength of LA-GAGR-T15 gel beads was stronger, and the strain was less likely to leak out. The average D-LA production was (72.90±2.79) g/L after fermentation for ten recycles (720 h) using LA-GAGR-T15 gel beads as the starting strain and glucose as the substrate, which was 33.85% higher than that of calcium alginate-T15 gel beads and 37.70% higher than that of free T15. Subsequently, glucose was replaced by enzymatically hydrolyzed corn straw and fermented for ten recycles (240 h) using LA-GAGR-T15 gel beads. The yield of D-LA reached (1.74±0.79) g/(L·h), which was much higher than that of using free bacteria. The wear rate of gel beads was less than 5% after ten recycles, which indicated that LA-GAGR is a good carrier for cell immobilization and can be widely used in industrial fermentation. This study provides basic data for the industrial production of D-LA using cell-recycled fermentation, and provides a new way for the biorefinery of D-LA from corn straw.
Fermentation
;
Lactobacillus delbrueckii
;
Zea mays
;
Lactic Acid
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Alginates/chemistry*
;
Glucose
2.Molecular characterization and efficacy evaluation of a transgenic corn event for insect resistance and glyphosate tolerance.
Miao-Miao LIU ; Xiao-Jing ZHANG ; Yan GAO ; Zhi-Cheng SHEN ; Chao-Yang LIN
Journal of Zhejiang University. Science. B 2018;19(8):610-619
A transgenic maize event ZD12-6 expressing a Bacillus thuringiensis (Bt) fusion protein Cry1Ab/Cry2Aj and a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein G10 was characterized and evaluated. Southern blot analysis indicated that ZD12-6 is a single copy integration event. The insert site was determined to be at chromosome 1 by border sequence analysis. Expression analyses of Bt fusion protein Cry1Ab/Cry2Aj and the EPSPS protein G10 suggested that they are both expressed stably in different generations. Insect bioassays demonstrated that the transgenic plants are highly resistant to Asian corn borer (Ostrinia furnacalis), cotton boll worm (Helicoverpa armigera), and armyworm (Mythimna separata). This study suggested that ZD12-6 has the potential to be developed into a commercial transgenic line.
3-Phosphoshikimate 1-Carboxyvinyltransferase/metabolism*
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Animals
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Bacillus thuringiensis Toxins
;
Bacterial Proteins/metabolism*
;
China
;
Disease Resistance/genetics*
;
Drug Resistance/genetics*
;
Endotoxins/metabolism*
;
Gene Expression Profiling
;
Glycine/chemistry*
;
Hemolysin Proteins/metabolism*
;
Insecta
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Plant Diseases/prevention & control*
;
Plants, Genetically Modified/genetics*
;
Zea mays/genetics*
;
Glyphosate
3.A corn straw-based diet increases release of inflammatory cytokines in peripheral blood mononuclear cells of dairy cows.
Yan-Yi CHE ; Xiao-Jing XIA ; Bo-Ping HE ; Yuan-Yuan GAO ; Wen-Bo REN ; Hong-Tao LIU ; Jian-Fang LIU ; Ting-Hao HUANG ; Wen-Yu HAN ; Lian-Cheng LEI
Journal of Zhejiang University. Science. B 2018;19(10):796-806
Recent studies have shown that diet can affect the body's immunity. Roughage of dairy cows consists of a variety of plant materials which make different contributions to health. This study investigated the effect of different roughages on the immunity of dairy cows. Serum, peripheral blood mononuclear cells (PBMCs), and milk samples were collected from 20 multiparous mid-lactation cows fed mixed forage (MF)- or corn straw (CS)-based diets. Expression profile analysis was used to detect the differentially expressed genes (DEGs) from PBMCs. The results showed that milk protein in the MF group increased to 3.22 g/100 ml, while that of the CS group milk was 2.96 g/100 ml; by RNA sequencing, it was found that 1615 genes were differentially expressed between the CS group and the MF group among the 24 027 analyzed probes. Gene ontology (GO) and pathway analysis of DEGs suggested that these genes (especially genes coding cytokines, chemokine and its receptors) are involved in the immune response. Results were confirmed at the protein level via detecting the levels of interleukin-2 (IL-2), IL-6, IL-10, IL-12, leptin (LEP), interferon-γ (IFN-γ), transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α) in peripheral blood by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay analysis. Our data supported the conclusions that the protein content in milk of the MF group was higher than that of the CS group, the CS-based diets induced more release of cytokines than the MF-based diets in dairy cows' PBMCs, and milk protein content may be affected by cytokines.
Animals
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Cattle/immunology*
;
Cytokines/physiology*
;
Diet
;
Female
;
Gene Ontology
;
Leukocytes, Mononuclear/immunology*
;
Milk/chemistry*
;
Transforming Growth Factor beta/physiology*
;
Zea mays
4.Global proteomic and phosphoproteomic analysis of the premature maize anther.
Zhimin ZHANG ; Juanying YE ; Haifei LONG ; Yue HONG ; Pingli LU
Chinese Journal of Biotechnology 2016;32(7):937-955
Reversible phosphorylation plays a crucial role in regulating protein activities and functions. Sexual reproduction directly affects yield of most agricultural crops. As the male reproductive organ, anther generates microspores (pollen), delivering gametes (sperms) to complete double fertilization in higher plants. Here, we took the advantage of Nano UHPLC-MS/MS to analyze maize (Zea mays, B73) early anthers at proteomic and phosphoproteomic levels, to explore the protein and phosphorylation modification regulatory networks controlling maize anther development. Our proteomic analysis identified 3 016 unique peptides, belonging to 1 032 maize proteins. MapMan analysis revealed variously potential proteins associated with maize anther development, such as receptor-like kinases (GRMZM2G082823_P01 and GRMZM5G805485_P01). Using phospho-peptides enriched by TiO2 affinity chromatography, our phosphoproteomic analysis detected 257 phospho-peptides from 210 phosphoproteins, discovering 223 phosphosites. Compared to the 86 maize phosphoproteins collected in the Plant Protein Phosphorylation Data Base (P3DB), we found that 203 phosphoproteins and 218 phosphosites were not revealed before. Further bioinformatics analysis revealed that phosphorylation of 14-3-3 proteins, kinases, phosphatases, transcription factors, cell cycle and chromatin structure related proteins might play important roles in regulating normal anther development in maize. Our findings not only enlarged the maize phosphoproteome data, but also provided information for analyzing the molecular mechanism controlling maize anther development at genetic and biochemical levels.
Crops, Agricultural
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chemistry
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Phosphoproteins
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chemistry
;
Phosphorylation
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Plant Proteins
;
chemistry
;
Pollen
;
chemistry
;
Proteome
;
Tandem Mass Spectrometry
;
Zea mays
;
chemistry
5.Surface display of phytase on Saccharomyces cerevisiae for efficient bioethanol production from corn starch.
Yan XIAO ; Xianzhong CHEN ; Wei SHEN ; Haiquan YANG ; You FAN
Chinese Journal of Biotechnology 2015;31(12):1700-1710
Production of bioethanol using starch as raw material has become a very prominent technology. However, phytate in the raw material not only decreases ethanol production efficiency, but also increases phosphorus discharge. In this study, to decrease phytate content in an ethanol fermentationprocess, Saccharomyces cerevisiae was engineered forheterologous expression of phytase on the cell surface. The phy gene encoding phytase gene was fused with the C-terminal-half region of α-agglutinin and then inserted downstream of the secretion signal gene, to produce a yeast surface-display expression vector pMGK-AG-phy, which was then transformed into S. cerevisiae. The recombinant yeast strain, PHY, successfully displayed phytase on the surface of cells producing 6.4 U/g wet cells and its properties were further characterized. The growthrate and ethanol production of the PHY strain were faster than the parent S. cerevisiae strain in the fermentation medium by simultaneous saccharification and fermentation. Moreover, the phytate concentration decreased by 91% in dry vinasse compared to the control. In summary, we constructed recombinant S. cerevisiae strain displaying phytase on the cell surface, which could effectively reduce the content of phytate, improve the utilization value of vinasse and reduce the discharge of phosphorus. The strain reported here represents a useful novel engineering platform for developing an environment-friendly system for bioethanol production from a corn substrate.
6-Phytase
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metabolism
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Biofuels
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Ethanol
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chemistry
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Fermentation
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Industrial Microbiology
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Saccharomyces cerevisiae
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metabolism
;
Starch
;
chemistry
;
Zea mays
;
chemistry
6.Biomass fast pyrolysis for bio-oil production in a fluidized bed reactor under hot flue atmosphere.
Ning LI ; Xiang WANG ; Xueyuan BAI ; Zhihe LI ; Ying ZHANG
Chinese Journal of Biotechnology 2015;31(10):1501-1511
Fast pyrolysis experiments of corn stalk were performed to investigate the optimal pyrolysis conditions of temperature and bed material for maximum bio-oil production under flue gas atmosphere. Under the optimized pyrolysis conditions, furfural residue, xylose residue and kelp seaweed were pyrolyzed to examine their yield distributions of products, and the physical characteristics of bio-oil were studied. The best flow rate of the flue gas at selected temperature is obtained, and the pyrolysis temperature at 500 degrees C and dolomite as bed material could give a maximum bio-oil yield. The highest bio-oil yield of 43.3% (W/W) was achieved from corn stalk under the optimal conditions. Two main fractions were recovered from the stratified bio-oils: light oils and heavy oils. The physical properties of heavy oils from all feedstocks varied little. The calorific values of heavy oils were much higher than that of light oils. The pyrolysis gas could be used as a gaseous fuel due to a relatively high calorific value of 6.5-8.5 MJ/m3.
Biofuels
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Biomass
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Bioreactors
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Furaldehyde
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chemistry
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Hot Temperature
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Kelp
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Temperature
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Xylose
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chemistry
;
Zea mays
7.Development of an aptamer/fluorescence dye PicoGreen-based method for detection of fumonisin B1.
Hailuan GUI ; Qingri JIN ; Yajun ZHANG ; Xiaodu WANG ; Yongchun YANG ; Chunyan SHAO ; Changyong CHENG ; Fangfang WEI ; Yang YANG ; Menghua YANG ; Houhui SONG
Chinese Journal of Biotechnology 2015;31(9):1393-1400
Fumonisin B1 (FB1) is a carcinogenic mycotoxin found in commodities such as corn and corn-originated products. An aptamer-based method for detection of FB1 was developed using the fluorescent dye PicoGreen, which can recognize and bind double-stranded DNA. A peak fluorescence of PicoGreen was obtained in 15 min in the presence of FB1 aptamer, which formed a double-stranded hybridizer DNA with its complementary strand. The excitation and emission wavelengths for PicoGreen detection were 480 nm and 520 nm, respectively. The sensitivity of this aptamer/PicoGreen-based method was 0.1 μg/L. This method showed a good linearity for FB1 concentration ranging from 0.1 to 1 μg/L. The entire detection procedure for FB1 could be completed within 40 min. No cross reactions were observed with any other mycotoxins against aflatoxin B1, ochratoxin A, citrinin and zearalenone, demonstrating high specificity towards FB1 aptamer. Agreement between commercial, antibody-based enzyme-linked immunosorbent assay (ELISA) kit and aptamer method was excellent with a kappa value of 0.857. Taken together, this aptamer/PicoGreen-based method is more cost-effective, time-saving and useful than ELISA for detection of FB1.
Aflatoxin B1
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Enzyme-Linked Immunosorbent Assay
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Fluorescence
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Fluorescent Dyes
;
chemistry
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Fumonisins
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analysis
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Mycotoxins
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analysis
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Ochratoxins
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Organic Chemicals
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chemistry
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Staining and Labeling
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Zea mays
8.Antioxidant and antigenotoxic activity of bioactive extracts from corn tassel.
Li-chun WANG ; Ya-qin YU ; Min FANG ; Cai-gui ZHAN ; Hong-yan PAN ; Yong-ning WU ; Zhi-yong GONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(1):131-136
This study is designed to evaluate antioxidant and antigenotoxic activities of corn tassel extracts (CTTs). The major bioactive components of CTTs include flavonoid, saponin and polysaccharide. The antioxidant properties of the three bioactive components of CTTs were investigated by Ferric Reducing Antioxidant Property (FRAP) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assays. The activities of the extracts were determined by assessing the inhibition of mutagenicity of the direct-acting mutagen fenaminosulf, sodium azide, and indirect-acting mutagen 2-aminofluorene using the Ames test (strains TA98 and TA100). The results showed that the extraction rates of flavonoid, saponin, and polysaccharide from the dried corn tassels were 1.67%, 2.41% and 4.76% respectively. DPPH and FRAP assay strongly demonstrated that CTTs had antioxidant properties. CTTs at doses of 625, 1250 and 2500 μg per plate reduced 2-aminofluorene mutagenicity by 12.52%, 28.76% and 36.49% in Salmonella typhimurium TA98 strain assay respectively and by 10.98%, 25.27% and 37.83%, at the same doses in Salmonella typhimurium TA100 assay system, respectively. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay showed that the different concentrations of CTTs inhibited the proliferation of MGC80-3 cells in a dose-dependent manner (P<0.01). It is concluded that these integrated approaches to antioxidant and antigenotoxicity assessment may be useful to study corn tassel as a natural herbal material.
Antimutagenic Agents
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pharmacology
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Antioxidants
;
pharmacology
;
Biphenyl Compounds
;
antagonists & inhibitors
;
metabolism
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Cell Line, Tumor
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Cell Survival
;
drug effects
;
Flavonoids
;
pharmacology
;
Fluorenes
;
pharmacology
;
Free Radical Scavengers
;
pharmacology
;
Humans
;
Inflorescence
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chemistry
;
Mutagens
;
pharmacology
;
Picrates
;
antagonists & inhibitors
;
metabolism
;
Plant Extracts
;
pharmacology
;
Polysaccharides
;
pharmacology
;
Salmonella typhimurium
;
drug effects
;
genetics
;
Saponins
;
pharmacology
;
Zea mays
;
chemistry
9.Allelopathic effects of aqueous extracts from Panax notoginseng on three maize varieties (Zea mays).
Zi-Long ZHANG ; Jun-Ling HOU ; Wen-Quan WANG ; Zhi-Xin ZHANG ; Shi-Xiu ZHANG
China Journal of Chinese Materia Medica 2014;39(4):594-600
It has been showed that there were obvious obstacle effects of Panax notoginseng replanting. Crop rotation was the main effective technique to overcome the obstacle. To find a reasonable crop rotation system for P. notoginseng, aqueous extracts from root, stem and leaf of P. notoginseng were analyzed for allelopathic effect on three maize varieties (which are often grown in regions where P. notoginseng grown). The main results were as follows: (1) Allelopathic effect of P. notoginseng stem and leaf extracts on the three other tested plants was stronger than that of root extracts; (2) Corn was more vulnerable to the effects of allelochemicals at seedling stage than at germination stage, and the corn root was more sensitive than aerial part to allelochemicals; (3) Lusan No. 3 and Yunrui No. 1 showed resistance to P. notoginseng allelopathy, with respective comprehensive sensitivity indexes (M3) of - 0.089 3 and -0.159 2, while Bainuo No. 1 is sensitive at M3 = -0.261 0. It then can be concluded that Lusan No. 3 and Yunrui No. 1 may be an alternative rotation plants for overcoming P. notoginseng continuous cropping obstacle.
Allelopathy
;
Panax notoginseng
;
chemistry
;
growth & development
;
Pheromones
;
pharmacology
;
Plant Extracts
;
pharmacology
;
Plant Leaves
;
chemistry
;
Plant Roots
;
chemistry
;
Plant Stems
;
chemistry
;
Zea mays
;
drug effects
;
growth & development
10.Phenolic foam prepared by lignin from a steam-explosion derived biorefinery of corn stalk.
Chinese Journal of Biotechnology 2014;30(6):901-910
To increase the integral economic effectiveness, biorefineries of lignocellulosic materials should not only utilize carbohydrates hydrolyzed from cellulose and hemicellulose but also use lignin. We used steam-exploded corn stalk as raw materials and optimized the temperature and alkali concentration in the lignin extraction process to obtain lignin liquor with higher yield and purity. Then the concentrated lignin liquor was used directly to substitute phenol for phenolic foam preparation and the performances of phenolic foam were characterized by microscopic structure analysis, FTIR, compression strength and thermal conductivity detection. The results indicated that, when steam-exploded corn stalk was extracted at 120 degrees C for 2 h by 1% NaOH with a solid to liquid ratio of 1:10, the extraction yield of lignin was 79.67%. The phenolic foam prepared from the concentrated lignin liquor showed higher apparent density and compression strength with the increasing substitution rate of lignin liquor. However, there were not significant differences of thermal conductivity and flame retardant properties by the addition of lignin, which meant that the phenolic foam substituted by lignin liquor was approved for commercial application. This study, which uses alkali-extracted lignin liquor directly for phenolic foam preparation, provides a relatively simple way for utilization of lignin and finally increases the overall commercial operability ofa lignocellulosic biorefinery derived by steam explosion.
Biotechnology
;
methods
;
Chemical Fractionation
;
Hot Temperature
;
Lignin
;
chemistry
;
Phenols
;
chemistry
;
Plant Stems
;
chemistry
;
Steam
;
Zea mays
;
chemistry

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