1.High titer ethanol production from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw.
Liang WANG ; Jianquan LIU ; Zhe ZHANG ; Feiyang ZHANG ; Junli REN ; Fubao SUN ; Zhenyu ZHANG ; Cancan DING ; Qiaowen LIN
Chinese Journal of Biotechnology 2015;31(10):1468-1483
The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels
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Carbohydrates
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chemistry
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Cellulose
;
chemistry
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Ethanol
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metabolism
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Fermentation
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Furaldehyde
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chemistry
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Glycerol
;
chemistry
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Hydrolysis
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Triticum
2.Progress and application prospects of glutamine synthase in plants.
Wanjun FENG ; Guofang XING ; Xulong NIU ; Chen DOU ; Yuanhuai HAN
Chinese Journal of Biotechnology 2015;31(9):1301-1312
Nitrogen is one of the most important nutrient elements for plants and a major limiting factor in plant growth and crop productivity. Glutamine synthase (GS) is a key enzyme involved in the nitrogen assimilation and recycling in plants. So far, members of the glutamine synthase gene family have been characterized in many plants such as Arabidopsis, rice, wheat, and maize. Reports show that GS are involved in the growth and development of plants, in particular its role in seed production. However, the outcome has generally been inconsistent, which are probably derived from the transcriptional and post-translational regulation of GS genes. In this review, we outlined studies on GS gene classification, QTL mapping, the relationship between GS genes and plant growth with nitrogen and the distribution characters, the biological functions of GS genes, as well as expression control at different regulation levels. In addition, we summarized the application prospects of glutamine synthetase genes in enhancing plant growth and yield by improving the nitrogen use efficiency. The prospects were presented on the improvement of nitrogen utility efficiency in crops and plant nitrogen status diagnosis on the basis of glutamine synthase gene regulation.
Arabidopsis
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Genes, Plant
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Glutamate-Ammonia Ligase
;
genetics
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Nitrogen
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metabolism
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Oryza
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Plants
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enzymology
;
genetics
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Triticum
;
Zea mays
3.A kinetic approach to evaluate salinity effects on carbon mineralization in a plant residue-amended soil.
Farshid NOURBAKHSH ; Ahmad R SHEIKH-HOSSEINI
Journal of Zhejiang University. Science. B 2006;7(10):788-793
The interaction of salinity stress and plant residue quality on C mineralization kinetics in soil is not well understood. A laboratory experiment was conducted to study the effects of salinity stress on C mineralization kinetics in a soil amended with alfalfa, wheat and corn residues. A factorial combination of two salinity levels (0.97 and 18.2 dS/m) and four levels of plant residues (control, alfalfa, wheat and corn) with three replications was performed. A first order kinetic model was used to describe the C mineralization and to calculate the potentially mineralizable C. The CO(2)-C evolved under non-saline condition, ranged from 814.6 to 4842.4 mg CO(2)-C/kg in control and alfalfa residue-amended soils, respectively. Salinization reduced the rates of CO(2) evolution by 18.7%, 6.2% and 5.2% in alfalfa, wheat and corn residue-amended soils, respectively. Potentially mineralizable C (C(0)) was reduced significantly in salinized alfalfa residue-treated soils whereas, no significant difference was observed for control treatments as well as wheat and corn residue-treated soils. We concluded that the response pattern of C mineralization to salinity stress depended on the plant residue quality and duration of incubation.
Carbon
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chemistry
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Carbon Dioxide
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chemistry
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Cellulose
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metabolism
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Ecosystem
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Kinetics
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Medicago sativa
;
metabolism
;
Models, Chemical
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Plants
;
metabolism
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Salts
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chemistry
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pharmacology
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Soil
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Soil Pollutants
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Triticum
;
metabolism
;
Zea mays
;
metabolism
4.Nutritional composition of Pakistani wheat varieties.
Journal of Zhejiang University. Science. B 2007;8(8):555-559
Pakistani wheat varieties are grown over a wide agro-climatic range and as such are anticipated to exhibit yield and quality differences. It is therefore necessary to investigate the nutritional status of wheat varieties in terms of biochemical and physiochemical characteristics available for food and nutritional purposes in Pakistan. The result shows that wheat grains of different varieties contain a net protein level of 9.15%-10.27%, 2.15%-2.55% total fats, 1.72%-1.85% dietary fibers, 77.65x10(-6)-84.25x10(-6) of potassium and 7.70x10(-6)-35.90x10(-6) of sodium ions concentration, 0.24x10(-6)-0.84x10(-6) of phosphorus, 1.44%-2.10% ash, 31.108-43.602 g of thousand grain mass (TGM) and 8.38%-9.67% moisture contents. This study is significant in providing an opportunity to explore the available wheat varieties and to further improve their nutritional excellence and also essential for setting nutritional regulations for domestic and export purposes.
Chemical Phenomena
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Chemistry, Physical
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Dietary Fiber
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metabolism
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Fats
;
metabolism
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Minerals
;
metabolism
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Nutritive Value
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Pakistan
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Plant Proteins
;
metabolism
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Triticum
;
chemistry
;
classification
;
metabolism
5.Expression of peaT1 gene from Alternaria tenuissima in Pichia pastoris and its function.
Yanfeng LIU ; Hongmei ZENG ; Shanjiang YU ; Xiufen YANG ; Jianjun MAO ; Dewen QIU
Chinese Journal of Biotechnology 2009;25(3):413-417
In this study, peaT1 gene was subcloned into the Pichia pastoris expression vector pPIC9K, which contained both the methanol-inducible promoter and the transcription terminator of the AOX1 gene, resulting the plasmid pPIC9K-peaT1. The recombinant plasmid was linearized by Sal I or Bgl II and transformed into P. pastoris GS115 by electroporation method. Recombinant strain was screened by Minimal Dextrose Medium and further confirmed by PCR. The gene was in frame integrated into the Pichia genome through homologous recombination, resulting the recombinant strain. Regulated by the alpha-Factor, promoter of AOX1 gene and termination signal of yeast genomic, the recombinant protein was expressed and secreted into the supernatant. The SDS-PAGE analysis indicated that the apparent molecular weight of target protein was about 35 kD. Bioassay results showed that the inhibition rate of the expressed protein against TMV was 30.37%. The supernatant was collected and then purified by anion exchange chromatography. This protein can promote seedling growth of wheat obviously.
Alternaria
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genetics
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Fungal Proteins
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biosynthesis
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genetics
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pharmacology
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Pichia
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genetics
;
metabolism
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Recombinant Proteins
;
biosynthesis
;
genetics
;
pharmacology
;
Triticum
;
drug effects
;
growth & development
6.A modified TAIL-PCR and its application in isolating gene promoter of wheat.
Yanguang QIU ; Jinghan TIAN ; Rongchao GE ; Baocun ZHAO ; Yinzhu SHEN ; Zhanjing HUANG
Chinese Journal of Biotechnology 2008;24(4):695-699
Using a modified TAIL-PCR technique, the 5' -flanking region of the X gene in wheat was successfully isolated. Two novel modifications of the TAIL-PCR were introduced here: using a battery of random 10-mers as the short arbitrary primers instead of three degenerate 16-mers; using 29 degrees C instead of 44 degrees C as the annealing temperature for the low-stringency cycle; increasing five high-stringency cycles and reducing five low-stringency cycles; and using single primers for the third round of product identification. Isolated 5' -flanking region was fused to the GUS gene, and tested for expression in Arabidopsis plants. Histochemical analysis of the transgenic plants showed the report gene was driven by isolated 5'-flanking region. Modified TAIL-PCR technique could isolate rapidly the promoter of any gene from organisms with large genomes.
Base Sequence
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Genes, Plant
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genetics
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Molecular Sequence Data
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Polymerase Chain Reaction
;
methods
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Promoter Regions, Genetic
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genetics
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Triticum
;
genetics
;
metabolism
7.Sequencing and expression analysis of a novel HMW-glutenin gene Kx from Crithopsis delileana.
Zhi-Fu GUO ; Ze-Hong YAN ; Yu-Ming WEI ; You-Liang ZHENG
Chinese Journal of Biotechnology 2005;21(3):375-379
Using SDS-PAGE analysis, the High-molecular-weight (HMW) glutenin subunits of two Crithopsis delileana accessions were detected. It was found that the two accessions had the same HMW glutenin subunits. Only one HMW glutenin subunit with the similar electrophoresis mobility to the y-type HMW glutenin subunit of hexaploid wheat was observed in C. delileana. However, It was indicated that this glutenin subunit was an x-type glutenin subunit Kx by gene sequence analysis. The full coding region of Kx gene is 2052bp and could encodes a mature protein with 661 amino acid residues. The Kx gene could be expressed in the bacterial expression system, and the expressed protein had the same electrophoresis mobility as that in the seed of C. delileana. The primary structure of Kx subunit was very similar to the x-type HMW glutenin subunits encoded by the A, B and D genomes of wheat, the C and U genomes of Aegilops, and the R genome of Secale cereale. In the phylogenetic analysis, Kx subunit was clustered together with Ax1 subunit by an interior parallelled branch. In conclsion, Kx gene is a novel x type glutenin subunit gene from C. delileana.
Amino Acid Sequence
;
Base Sequence
;
Genes, Plant
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Glutens
;
genetics
;
metabolism
;
Molecular Sequence Data
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Plant Proteins
;
genetics
;
metabolism
;
Poaceae
;
genetics
;
metabolism
;
Triticum
;
genetics
;
metabolism
8.Laboratory simulated dissipation of metsulfuron methyl and chlorimuron ethyl in soils and their residual fate in rice, wheat and soybean at harvest.
Nilanjan SANYAL ; Sukhendu Kumar PRAMANIK ; Raktim PAL ; Ashim CHOWDHURY
Journal of Zhejiang University. Science. B 2006;7(3):202-208
Two sulfonylurea herbicides, metsulfuron methyl (Ally 20 WP) and chlorimuron ethyl (Classic 25 WP) were evaluated for their dissipation behaviour in alluvial, coastal saline and laterite soils under laboratory incubated condition at 60% water holding capacity of soils and 30 degrees C temperature was maintained. In field study herbicides were applied twice for the control of grasses, annual and perennials broad leaves weeds and sedges in rice, wheat and soybean to find out the residual fate of both the herbicides on different matrices of respective crops after harvest. Extraction and clean up methodologies for the herbicides were standardized and subsequently analyzed by HPLC. The study revealed that the half-lives of metsulfuron methyl and chlorimuron ethyl ranged from 10.75 to 13.94 d irrespective of soils and doses applied. Field trials with rice, wheat and soybean also revealed that these two herbicides could safely be recommended for application as no residues were detected in the harvest samples.
Arylsulfonates
;
analysis
;
pharmacokinetics
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Herbicides
;
analysis
;
pharmacokinetics
;
Oryza
;
metabolism
;
Pyrimidines
;
analysis
;
pharmacokinetics
;
Soil
;
analysis
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Soybeans
;
metabolism
;
Sulfonylurea Compounds
;
analysis
;
pharmacokinetics
;
Triticum
;
metabolism
9.Molecular and biochemical analysis of two genes encoding dehydroascorbate reductase in common wheat.
Chunmei YU ; Yanping YANG ; Xinyan LIU ; Rong ZHOU ; Liang HUA ; He WEI ; Shengjie DING ; Daowen WANG
Chinese Journal of Biotechnology 2009;25(10):1483-1489
Dehydroascorbate reductase (DHAR) plays an important role in the recycling of ascorbic acid. In this work, we isolated the full length cDNA clones of two different DHAR genes (tentatively named as TaDHAR1 and TaDHAR2, respectively) from common wheat. Semi-quantitative PCR experiments showed that TaDHAR1 and TaDHAR2 were transcribed in many vegetative and reproductive organs examined in this work. Transient expression analysis using wheat protoplasts indicated that the protein products of TaDHAR1 and TaDHAR2 may be located in the cytoplasm. The cDNAs of TaDHAR1 and TaDHAR2 were expressed in the bacterial cells, and resultant histidine tagged recombinant proteins could be efficiently purified using nickel chelate affinity chromatography. In vitro enzyme activity assays revealed that the recombinant TaDHAR1 and TaDHAR2 proteins could all convert dehydroascorbate (DHA) to AsA. The two proteins exhibited higher activity levels at 37 degrees C than at 25 degrees C. Under the two temperature conditions, the optimal pH for TaDHAR1 and TaDHAR2 was both around 7.5. The major difference between TaDHAR1 and TaDHAR2 is the activity under pH 6.0 and 7.0 at 25 degrees C. The results and resources obtained in this study may be useful for further research into the physiological role of TaDHAR genes in AsA metabolism in crop plants under normal or stressed conditions.
Amino Acid Sequence
;
Cloning, Molecular
;
Genes, Plant
;
genetics
;
Molecular Sequence Data
;
Oxidoreductases
;
genetics
;
metabolism
;
Plant Proteins
;
genetics
;
metabolism
;
Recombinant Proteins
;
genetics
;
metabolism
;
Triticum
;
enzymology
;
genetics
10.Detection, isolation, and characterization of a novel impurity from several folic acid products.
Qirong SHEN ; Quan HE ; Yuanjiang PAN ; Cuirong SUN
Journal of Zhejiang University. Science. B 2022;23(7):597-606
Folic acid belongs to the group of water-soluble B vitamins and naturally exists in multiple forms in a wide variety of foods such as legumes, vegetables, liver, and milk (Iyer and Tomar, 2009; Lyon et al., 2020). It is involved in many biochemical reactions critical for cell division, such as purine and pyrimidine biosynthesis, DNA/RNA biosynthesis, and amino acid metabolism (Iyer and Tomar, 2009). Mammals cannot synthesize folic acid and thus they must acquire it from food. Although folic acid is ubiquitous in foods, folic acid deficiency still often occurs due to various causes such as unhealthy diet (Hildebrand et al., 2021; Iimura et al., 2022), disease-related malabsorption (Arcot and Shrestha, 2005), medication-related depletion (Arcot and Shrestha, 2005), or vitamin B12 deficiency (Fishman et al., 2000). Folic acid deficiency has been associated with several health problems, such as anemia (Carmel, 2005; Bailey and Caudill, 2012), cancer (Duthie, 1999), cardiovascular diseases (Wald et al., 2002), neural tube defects in newborns (van der Put et al., 2001), neuropsychiatric dysfunction (Shea et al., 2002), depression (Falade et al., 2021), inflammatory diseases (Suzuki and Kunisawa, 2015; Jones et al., 2019), and eye diseases (Sijilmassi, 2019). To prevent folic acid deficiency, its daily intake (400 μg/d) has been recommended for adults in the European Union, and its increased intake (600 μg/d) is advised for women before and during pregnancy (FAO/WHO, 2002; IOM, 2004). The New Zealand government mandated the fortification of non-organic wheat flour with folic acid in July 2021, and the UK government mandated the fortification of non-wholemeal wheat flour with folic acid in September 2021 (Haggarty, 2021).
Adult
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Animals
;
Female
;
Flour
;
Folic Acid/metabolism*
;
Folic Acid Deficiency/prevention & control*
;
Food, Fortified
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Humans
;
Infant, Newborn
;
Mammals/metabolism*
;
Pregnancy
;
Triticum/metabolism*