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1.
《Process Biochemistry》2007,42(3):335-343
A comparative study of the performance of two types of adsorbent (Streamline Quartz Base and Upfront Matrices), derivatized with the same affinity ligand (RPAP) to recover C595 diabody fragment (dbFv) from Escherichia coli lysates has been undertaken. Both streamline and Upfront Matrices are characterized by a particle size range of 100–300 μm. Streamline has a density of 1.20 g cm−3 and ligand concentration of 0.85 μmol ml−1. Upfront has a density of 1.35 g cm−3 and ligand concentration of 0.83 μmol ml−1. The release of C595 dbFv from E. coli cells was achieved by a chemical lysis method. The recovery performance of both adsorbents was evaluated in terms of operational productivity and elution yield of C595 dbFv in packed bed (clarified feedstock) and expanded bed (unclarified and clarified feedstock) chromatography systems. Streamline and Upfront adsorbents exhibited diabody operational productivities of 131 and 202 mg l−1, respectively, with an elution yield of 92 and 94%, respectively, in packed bed operation. Streamline and Upfront adsorbents exhibited diabody operational productivities of 54.5 and 123.7 mg l−1, respectively, with an elution yield of 89 and 92%, respectively, in expanded bed operation.  相似文献   

2.
The development of fermentation conditions for the production of C595 diabody fragment (dbFv) inE. coli HB2151 clone has been explored. Investigations were carried out to study the effect of carbon supplements over the expression period, the comparison of C595 dbfv production in synthetic and complex media, the influence of acetic acid upon antibody production, and comparison of one-stage and two-stage processes operated at batch or fed-batch modes in bioreactor. Yeast extract supplied during expression yielded more antibody fragment than any other carbon supply. The synthetic medium presented higher specific productivity (0.066 mg dbFv g−1 dry cell weight) when compared to the complex medium (0.044 mg dbFv g−1 DCW). The comparison of fermentation strategies demonstrated that (1) one-stage fed-batch fermentation performed higher C595 dbFv production than that operated in batch mode which was significantly affected by acetate concentration; (2) a two-stage batch operation could enhance C595 dbFv production. It was found that a concentration of 12.3 mg L−1 broth of C595 dbFv and a cell concentration of 10.8 g L−1 broth were achieved at the end of two-stage operation in 5-L fermentation.  相似文献   

3.
 Enzymatic hydrolysis of corncob and ethanol fermentation from cellulosic hydrolysate were investigated. After corncob was pretreated by 1% H2SO4 at 108 °C for 3 h, the cellulosic residue was hydrolyzed by cellulase from Trichoderma reesei ZU-02 and the hydrolysis yield was 67.5%. Poor cellobiase activity in T. reesei cellulase restricted the conversion of cellobiose to glucose, and the accumulation of cellobiose caused severe feedback inhibition to the activities of β-1,4-endoglucanase and β-1,4-exoglucanase in cellulase system. Supplementing cellobiase from Aspergillus niger ZU-07 greatly reduced the inhibitory effect caused by cellobiose, and the hydrolysis yield was improved to 83.9% with enhanced cellobiase activity of 6.5 CBU g−1 substrate. Fed-batch hydrolysis process was started with a batch hydrolysis containing 100 g l−1 substrate, with cellulosic residue added at 6 and 12 h twice to get a final substrate concentration of 200 g l−1. After 60 h of reaction, the reducing sugar concentration reached 116.3 g l−1 with a hydrolysis yield of 79.5%. Further fermentation of cellulosic hydrolysate containing 95.3 g l−1 glucose was performed using Saccharomyces cerevisiae 316, and 45.7 g l−1 ethanol was obtained within 18 h. The research results are meaningful in fuel ethanol production from agricultural residue instead of grain starch.  相似文献   

4.
Simultaneous saccharification and fermentation (SSF) of renewable cellulose for the production of 3-phenyllactic acid (PhLA) by recombinant Escherichia coli was investigated. Kraft pulp recovered from biomass fractionation processes was used as a model cellulosic feedstock and was hydrolyzed using 10–50 filter paper unit (FPU) g−1 kraft pulp of a commercial cellulase mixture, which increased the glucose yield from 21% to 72% in an enzyme dose-dependent manner. PhLA fermentation of the hydrolyzed kraft pulp by a recombinant E. coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens TK1 produced 1.9 mM PhLA. The PhLA yield obtained using separate hydrolysis and fermentation was enhanced from 5.8% to 42% by process integration into SSF of kraft pulp (20 g L−1) in a complex medium (pH 7.0) at 37 °C. The PhLA yield was negatively correlated with the initial glucose concentration, with a five-fold higher PhLA yield observed in culture medium containing 10 g L−1 glucose compared to 100 g L−1. Taken together, these results suggest that the PhLA yield from cellulose in kraft pulp can be improved by SSF under glucose-limited conditions.  相似文献   

5.
Conidiation and lytic enzyme production by Trichoderma viride at different solids concentration of pre-treated municipal wastewater sludge was examined in a 15-L fermenter. The maximum conidia concentration (5.94 × 107 CFU mL−1 at 96 h) was obtained at 30 g L−1 suspended solids. The maximum lytic enzyme activities were achieved around 12–30 h of fermentation. Bioassay against a fungal phytopathogen, Fusarium sp. showed maximum activity in the sample drawn around 96 h of fermentation at 30 g L−1 suspended solids concentration. Entomotoxicity against spruce budworm larvae showed maximum value ≈17290 SBU μL−1 at 30 g L−1 suspended solids concentration at the end of fermentation (96 h). Plant bioassay showed dual action of T. viride, i.e., disease prevention and growth promotion. The rheological analyses of fermentation sludges showed the pseudoplastic behaviour. In order to maintain required dissolved oxygen concentration ≥30%, the agitation and aeration requirements significantly increased at 35 g L−1 compared to 30 and 25 g L−1. The oxygen uptake rate and volumetric oxygen mass transfer coefficient, kLa at 35 g L−1 did not increase in comparison to 30 g L−1 due to rheological complexity of the broth during fermentation. Thus, the successful fermentation operation of the biocontrol fungus T. viride is a rational indication of its potential for mass-scale production for agriculture and forest sector as a biocontrol agent.  相似文献   

6.
The interest on use of lignocellulose for producing chemicals is increasing as these feedstocks are low cost, renewable and widespread sources of sugars. Corncob is an attractive raw material for xylitol production due to its high content of xylan. In this study, hemicellulose hydrolysate from corncobs without detoxification was used for xylitol production by Candida tropicalis CCTCC M2012462. Compared with prepared xylose medium, xylitol production with dilute acid hydrolysate medium does not seem to influence specific xylose reductase activity. The decrease in xylitol productivity with dilute acid hydrolysate medium is a result of a lower biomass concentration and lag-phase time. It appears that biomass growth rate is essential for xylitol production. In xylitol fermentation with a low initial inhibitors concentration and substrate feeding strategy, a maximal xylitol concentration of 38.8 g l−1 was obtained after 84 h of fermentation, giving a yield of 0.7 g g−1 xylose and a productivity of 0.46 g l−1 h−1.  相似文献   

7.
《Process Biochemistry》2007,42(4):740-744
The conversion of glycerol to 1,3-propanediol (PDO) using Klebsiella pneumoniae M5al under anaerobic condition was scaled up from scale 5 to 5000 l in series. A simple strategy for scale-up was to transfer the optimized conditions of a lab scale bioreactor to pilot-scale fermentation. Multistage inocula were developed and their fermentation abilities were assessed in a small-scale fermenter. The experimental results showed that inoculum development in the early steps of a scale-up process could influence the outcomes of a large scale fermentation. Through three-stage liquid inoculum development and a pulse addition of (NH4)2SO4 and yeast extract at 30 h of fermentation, the best results in a 5000 l fermentation were achieved leading to 58.8 g l−1 1,3-propanediol with a yield of 0.53 mol mol−1 glycerol and productivity of 0.92 g l−1 h−1. This is the first report on pilot-scale 1,3-propanediol production using K. pneumoniae.  相似文献   

8.
《Process Biochemistry》2007,42(3):352-362
The effects of medium components and environmental factors on the production of mycelial biomass and polysaccharide–peptide complexes (exobiopolymers) by Cordyceps sphecocephala J-201 were investigated in submerged cultures. The optimal temperature and initial pH for the production of both mycelial biomass and exobiopolymers in flask cultures were found to be 25 °C and pH 4–5, respectively. The optimal combination of the media constituents was as follows (g l−1): sucrose 40, yeast extract 6, polypepton 2, KH2PO4 0.46, K2HPO4 1, and MgSO4·7H2O 0.5. The results of bioreactor culture revealed that the maximum concentration of mycelial biomass (28.2 g l−1) was obtained at an agitation speed of 300 rpm and at an aeration rate of 2 vvm, whereas maximum exobiopolymer production (2.5 g l−1) was achieved at a milder agitation speed (150 rpm). There was a significant variance in mycelial morphology between different aeration conditions. Looser mycelial pellets were developed, and their size and hairiness increased as the aeration rate increased from 0.5 to 2.0 vvm, resulting in enhanced exobiopolymer production. The apparent viscosities of fermentation broth increased rapidly towards the end of fermentations at the conditions of high aeration rate and agitation speed, which were mainly due to high amount of mycelial biomass rather than exobiopolymers at the later stages of fermentation. The three different exobiopolymers (FR-I, -II, and -III) were fractionated by a gel filtration chromatography on Sepharose CL-6B. The carbohydrate and protein contents in each fraction were significantly different and the molecular weights of FR-I, FR-II, and FR-III were determined to be 1831, 27, and 2.2 kDa, respectively. The compositional analysis revealed that the three fractions of crude exobiopolymers consisted of acidic and nonpolar amino acids, such as aspartic acid, glutamic acid, glycine, and valine in protein moiety, and of mainly mannose and galactose in sugar moiety.  相似文献   

9.
《Process Biochemistry》2007,42(3):344-351
A strategy that optimization of medium compositions for maximum biomass followed by feeding of sucrose for maximum polysaccharide synthesis was developed for enhancing polysaccharide production in suspension culture of protocorm-like bodies (PLBs) of Dendrobium huoshanense C.Z. Tang et S.J. Cheng. In growth stage, the original half-strength MS medium was optimized with carbon sources, nitrogen sources and metal ion combinations. The effects of different carbon sources on PLBs growth were remarkable and sucrose at 35 g l−1 was the most suitable. Sole nitrate nitrogen of 30 mmol l−1 was the best for PLBs growth. Metal ions (Ca2+, Fe2+, Mn2+ and Zn2+) showed different influences on PLBs growth. The optimal concentration of Ca2+, Fe2+, Mn2+ and Zn2+ was 4.5 mmol l−1, 0.1 mmol l−1, 0.5 mmol l−1 and 0.06 mmol l−1, respectively. In the optimized medium (sucrose, nitrate, Ca2+, Fe2+, Mn2+ and Zn2+ concentration as described above, the other component concentration seen in half-strength MS), 33.9 g DW l−1 PLBs were harvested after 30 days of culture and biomass increase was improved 245% as compared with that in the original medium. In production stage, polysaccharide synthesis was significantly improved by the feeding sucrose. The maximum polysaccharide production (22 g l−1) was obtained in the case of 50 g l−1 sucrose feeding at day 30 of culture, which was about 109-fold higher than that in the original medium without feeding of sucrose.  相似文献   

10.
《Process Biochemistry》2007,42(1):112-117
A simple fed-batch process was developed using a modified variable specific growth rate feeding strategy for high cell density cultivation of Escherichia coli BL21 (DE3) expressing human interferon-gamma (hIFN-γ). The feeding rate was adjusted to achieve the maximum attainable specific growth rate during fed-batch cultivation. In this method, specific growth rate was changed from a maximum value of 0.55 h−1 at the beginning of feeding and then it was reduced to 0.4 h−1 at induction time.The final concentration of biomass and IFN-γ was reached to ∼115 g l−1 (DCW) and 42.5 g(hIFN-γ) l−1 after 16.5 h, also the final specific yield and overall productivity of recombinant hIFN-γ (rhIFN-γ) were obtained 0.37 g(hIFN-γ) g−1 DCW and 2.57 g(hIFN-γ) l−1 h−1, respectively. According to available data this is the highest specific yield and productivity that has been reported for recombinant proteins production yet.  相似文献   

11.
The capability of Corynebacterium glutamicum for glucose-based synthesis of itaconate was explored, which can serve as building block for production of polymers, chemicals, and fuels. C. glutamicum was highly tolerant to itaconate and did not metabolize it. Expression of the Aspergillus terreus CAD1 gene encoding cis-aconitate decarboxylase (CAD) in strain ATCC13032 led to the production of 1.4 mM itaconate in the stationary growth phase. Fusion of CAD with the Escherichia coli maltose-binding protein increased its activity and the itaconate titer more than two-fold. Nitrogen-limited growth conditions boosted CAD activity and itaconate titer about 10-fold to values of 1440 mU mg−1 and 30 mM. Reduction of isocitrate dehydrogenase activity via exchange of the ATG start codon to GTG or TTG resulted in maximal itaconate titers of 60 mM (7.8 g l−1), a molar yield of 0.4 mol mol−1, and a volumetric productivity of 2.1 mmol l−1 h−1.  相似文献   

12.
Spent coffee grounds (SCG) represent the main coffee industry residues with a great potential to be reutilized in various biotechnological processes. In this study, several carotenogenic yeasts strains were exploited for the production of vitamin-enriched biomass, cultivating in SCG-based media. The fermentation was firstly carried out in Erlenmeyer flasks in order to select the best biomass and pigment producer. Among four tested strains, Sporobolomyces roseus showed the highest potential for the accumulation of carotenoids. Maximum pigment concentration and yield was obtained when cultivating in SCG-based media, 12.59 mg l−1 and 1.26 mg g−1, respectively. Comparing both, the batch and the fed-batch cultivation modes, the strategy of sequential addition of pre-concentrated SCG media in the bioreactor gave higher biomass yield (maximum 41 g l−1 during 41–48 h after the beginning of fermentation). Thus, SCG can be considered as potentially promising industrial waste stream for economically feasible production of enriched yeasts biomass.  相似文献   

13.
Twenty-five aerobic phenol-degrading bacteria, isolated from different environmental samples on phenol agar after several subcultures in phenol broth, utilized phenol (0.2 g l−1) within 24 h, but removal of phenol was more rapid when other carbon sources were also present. A microtitre plate method was developed to determine growth rate, biofilm formation and respiratory activity of the strains isolated. Pseudomonas putida strains C5 and D6 showed maximum growth (as O.D. at 600 nm), P. putida D6 and unidentified bacterial strain M1 were more stable at high concentrations of phenol (0.8 g l−1), and P. putida C5 formed the greatest amount of biofilm in 0.5 g phenol l−1 medium. Measurement of dehydrogenase activity as reduction of triphenyl tetrazolium chloride supported data on growth rate and biofilm formation. The microtitre plate method provided a selective method for detection of the best phenol degrading and biofilm-forming microorganisms, and was also a rapid, convenient means of studying the effect of phenol concentration on growth rate and biofilm formation.  相似文献   

14.
In this work, straw hydrolysates were used to produce succinic acid by Actinobacillus succinogenes CGMCC1593 for the first time. Results indicated that both glucose and xylose in the straw hydrolysates were utilized in succinic acid production, and the hydrolysates of corn straw was better than that of rice or wheat straw in anaerobic fermentation of succinic acid. However, cell growth and succinic acid production were inhibited when the initial concentration of sugar, which was from corn straw hydrolysate (CSH), was higher than 60 g l?1. In batch fermentation, 45.5 g l?1 succinic acid concentration and 80.7% yield were attained after 48 h incubation with 58 g l?1 of initial sugar from corn straw hydrolysate in a 5-l stirred bioreactor. While in fed-batch fermentation, concentration of succinic acid achieved 53.2 g l?1 at a rate of 1.21 g l?1 h?1 after 44 h of fermentation. Our work suggested that corn straw could be utilized for the economical production of succinic acid by A. succinogenes.  相似文献   

15.
Cheese whey powder (CWP) solution with different CWP or sugar concentrations was fermented to ethanol in a continuous fermenter using pure culture of Kluyveromyces marxianus (DSMZ 7239). Sugar concentration of the feed CWP solution varied between 55 and 200 g l−1 while the hydraulic residence time (HRT) was kept constant at 54 h. Ethanol formation, sugar utilization and biomass formation were investigated as functions of the feed sugar concentration. Percent sugar utilization and biomass concentrations decreased and the effluent sugar concentration increased with increasing feed sugar concentrations especially for the feed sugar contents above 100 g l−1. Ethanol concentration and productivity (DP) increased with increasing feed sugar up to 100 g l−1 and then decreased with further increases in the feed sugar content. The highest ethanol concentration (3.7%, v v−1) and productivity (0.54 gE l−1 h−1) were obtained with the feed sugar content of 100 g l−1 or 125 g l−1. The ethanol yield coefficient (YP/S) was also maximum (0.49 gE gS−1) when the feed sugar was between 100 and 125 g l−1. The growth yield coefficient (YX/S) decreased steadily from 0.123 to 0.063 gX gS−1 when the feed sugar increased from 55 to 200 g l−1 due to adverse effects of high sugar contents on yeast growth. The optimal feed sugar concentration maximizing the ethanol productivity and sugar utilization was between 100 and 125 g l−1 under the specified experimental conditions.  相似文献   

16.
Succinate fermentation was investigated in Escherichia coli strains overexpressing cyanobacterium Anabaena sp. 7120 ecaA gene encoding carbonic anhydrase (CA). In strain BL21 (DE3) bearing ecaA, the activity of CA was 21.8 U mg−1 protein, whereas non-detectable CA activity was observed in the control strain. Meanwhile, the activity of phosphoenolpyruvate carboxylase (PEPC) increased from 0.2 U mg−1 protein to 1.13 U mg−1 protein. The recombinant bearing ecaA reached a succinate yield of 0.39 mol mol−1 glucose at the end of the fermentation. It was 2.1-fold higher than that of control strain which was just 0.19 mol mol−1 glucose. EcaA gene was also introduced into E. coli DC1515, which was deficient in glucose phosphotransferase, lactate dehydrogenase and pyruvate:formate lyase. Succinate yield can be further increased to 1.26 mol mol−1 glucose. It could be concluded that the enhancement of the supply of HCO3 in vivo by ecaA overexpression is an effective strategy for the improvement of succinate production in E. coli.  相似文献   

17.
《Process Biochemistry》2010,45(3):297-305
Pure glycerol and glycerol-rich product (GRP) obtained from the biodiesel industries were used as carbon source for the production of a new extracellular polysaccharide (EPS) by Pseudomonas oleovorans NRRL B-14682. The influence of temperature (20–40 °C) and pH (6.0–8.0) was studied. A temperature of 30 °C and pH control at 6.8 gave the maximum cell growth and EPS production. The culture attained a maximum cell dry weight (CDW) of 9.55 g l−1 and an EPS concentration of 11.82 g l−1 when cultivated with pure glycerol. GRP was a suitable carbon source, as shown by the slightly higher EPS concentration (12.18 g l−1). The EPS productivity obtained with GRP (3.85 g l−1 d−1) was almost twice that obtained with pure glycerol (2.00 g l−1 d−1). Also, the yield on glycerol was higher for the cultivation with GRP (0.36 g g−1) than for pure glycerol (0.28 g g−1). The EPS was a high molecular weight heteropolysaccharide, composed by neutral sugars (37–80 wt% galactose, 2–30 wt% glucose, 0.5–25 wt% mannose and 0.5–20 wt% rhamnose) and containing acyl group substituents (pyruvil, acetyl and succinyl were identified). The EPS forms highly viscous aqueous dispersions with many potential commercial applications.  相似文献   

18.
A thermo-alkaline pectate lyase (BliPelA) gene from an alkaliphilic Bacillus licheniformis strain was cloned and overexpressed in Escherichia coli. Mature BliPelA exhibited maximum activity at pH 11 and 70 °C, and demonstrated cleavage capability on a broad range of substrates such as polygalacturonic acid, pectins, and methylated pectins. The highest specific activity, of 320 U mg−1, was towards polygalacturonic acid. Significant ramie (Boehmeria nivea) fiber weight loss (21.5%) was obtained following enzyme treatment and combined enzyme-chemical treatment (29.3%), indicating a high ramie degumming efficiency of BliPelA. The total activity of recombinant BliPelA reached 1450.1 U ml−1 with a productivity of 48.3 U ml−1 h−1 under high-cell-density cultivation with a glycerol exponential feeding strategy for 30 h in 1-l fed-batch fermenter, and 1380.1 U ml−1 with a productivity of 57.5 U ml−1 h−1 after 24 h under constant glucose feeding in a 20-l fermenter using E. coli as the host. The enzyme yields reached 4.5 and 4.3 g l−1 in 1-l and 20-l fed-batch fermenters, respectively, which are higher than those of most reported alkaline Pels. Based on these promising properties and high-level production, BliPelA shows great potential for application in ramie degumming in textile industry.  相似文献   

19.
《Process Biochemistry》2007,42(6):1033-1038
Valienamine is an important medicinal intermediate with broad use in the synthesis of some stronger α-glucosidase inhibitors. In order to improve valienamine concentration in the fermentation broth and make the downstream treatment easy, a fed-batch process for the enhanced production of valienamine by Stenotrophomonas maltrophilia in a stirred tank bioreactor was developed. Results showed that supplementation of validamycin A in the process of cultivation could increase the valienamine concentration. One-pulse feeding was observed to be the best strategy. The maximum valienamine concentration of 2.35 g L−1 was obtained at 156 h when 86.4 g of validamycin A was added to a 15-L bioreactor containing 8 L fermentation medium with one-pulse feeding. The maximum valienamine concentration had a great improvement and was increased above 100% compared to batch fermentation in the stirred tank bioreactor. The pH-controlled experiments showed that controlling the pH in the process of one-pulse feeding fermentation had not obvious effect on the production of valienamine.  相似文献   

20.
There are currently few successful examples of using straw hemicellulose as a carbon source in the fermentation industry. In this paper, hemicellulose hydrolysates were recovered from steam-exploded wheat straw (SEWS) and used to produce microbial oil. The effects of the steam explosion treatment conditions, the elution temperature and the ratio of elution water to SEWS on sugar recovery were examined. A broth with 3.8 g l?1 of reducing sugar and 22.3 g l?1 of total soluble sugars was obtained with a 10-fold excess (w/w) of water at 40 °C to wash the SEWS treated under steam explosion conditions at 200 °C for 5 min. This broth was used to produce microbial oil by the oleaginous fungus Microsphaeropsis sp., which was able to secrete xylanase to degrade oligosaccharides from straw hemicellulose and accumulate microbial oil. Under optimized conditions, the oil concentration was 2.6 g l?1. The yield of oil from sugar consumed was 0.14 g g?1. The microbial oil produced by this research could be used as feedstock for biodiesel production because the microbial oil was primarily composed of neutral lipids. This research establishes a novel protocol for microbial oil production from straw hemicellulose.  相似文献   

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