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1.
BackgroundEthanol concentration (PE), ethanol productivity (QP) and sugar consumption (SC) are important values in industrial ethanol production. In this study, initial sugar and nitrogen (urea) concentrations in sweet sorghum stem juice (SSJ) were optimized for high PE (≥ 10%, v/v), QP, (≥ 2.5 g/L·h) and SC (≥ 90%) by Saccharomyces cerevisiae SSJKKU01. Then, repeated-batch fermentations under normal gravity (NG) and high gravity (HG) conditions were studied.ResultsThe initial sugar at 208 g/L and urea at 2.75 g/L were the optimum values to meet the criteria. At the initial yeast cell concentration of ~ 1 × 108 cells/mL, the PE, QP and SC were 97.06 g/L, 3.24 g/L·h and 95.43%, respectively. Repeated-batch fermentations showed that the ethanol production efficiency of eight successive cycles with and without aeration were not significantly different when the initial sugar of cycles 2 to 8 was under NG conditions (~ 140 g/L). Positive effects of aeration were observed when the initial sugar from cycle 2 was under HG conditions (180–200 g/L). The PE and QP under no aeration were consecutively lower from cycle 1 to cycle 6. Additionally, aeration affected ergosterol formation in yeast cell membrane at high ethanol concentrations, whereas trehalose content under all conditions was not different.ConclusionInitial sugar, sufficient nitrogen and appropriated aeration are necessary for promoting yeast growth and ethanol fermentation. The SSJ was successfully used as an ethanol production medium for a high level of ethanol production. Aeration was not essential for repeated-batch fermentation under NG conditions, but it was beneficial under HG conditions.How to cite: Sriputorn B, Laopaiboon P, Phukoetphim N, et al. Enhancement of ethanol production efficiency in repeated-batch fermentation from sweet sorghum stem juice: Effect of initial sugar, nitrogen and aeration. Electron J Biotechnol 2020;46. https://doi.org/10.1016/j.ejbt.2020.06.001  相似文献   

2.
BackgroundOptimization of nutrient feeding was developed to improve the growth of Bacillus subtilis in fed batch fermentation to increase the production of jiean-peptide (JAA). A central composite design (CCD) was used to obtain a model describing the relationship between glucose, total nitrogen, and the maximum cell dry weight in the culture broth with fed batch fermentation in a 5 L fermentor.ResultsThe results were analyzed using response surface methodology (RSM), and the optimized values of glucose and total nitrogen concentration were 30.70 g/L and 1.68 g/L in the culture, respectively. The highest cell dry weight was improved to 77.50 g/L in fed batch fermentation, which is 280% higher than the batch fermentation concentration (20.37 g/L). This led to a 44% increase of JAA production in fed batch fermentation as compared to the production of batch fermentation.ConclusionThe results of this work improve the present production of JAA and may be adopted for other objective products' production.  相似文献   

3.
BackgroundThe effect of diverse oxygen transfer coefficient on the l-erythrulose production from meso-erythritol by a newly isolated strain, Gluconobacter kondonii CGMCC8391 was investigated. In order to elucidate the effects of volumetric mass transfer coefficient (kLa) on the fermentations, baffled and unbaffled flask cultures, and fed-batch cultures were developed in present work.ResultsWith the increase of the kLa value in the fed-batch culture, l-erythrulose concentration, productivity and yield were significantly improved, while cell growth was not the best in the high kLa. Thus, a two-stage oxygen supply control strategy was proposed, aimed at achieving high concentration and high productivity of l-erythrulose. During the first 12 h, kLa was controlled at 40.28 h-1 to obtain high value for cell growth, subsequently kLa was controlled at 86.31 h-1 to allow for high l-erythrulose accumulation.ConclusionsUnder optimal conditions, the l-erythrulose concentration, productivity, yield and DCW reached 207.9 ± 7.78 g/L, 6.50 g/L/h, 0.94 g/g, 2.68 ± 0.17 g/L, respectively. At the end of fermentation, the l-erythrulose concentration and productivity were higher than those in the previous similar reports.  相似文献   

4.
BackgroundCurrently, microbial fermentation method has become the research hotspot for acetoin production. In our previous work, an acetoin-producing strain, Bacillus subtilis SF4-3, was isolated from Japanese traditional fermented food natto. However, its conversion of glucose to acetoin was relatively low. In order to achieve a high-efficient accumulation of acetoin in B. subtilis SF4-3, main medium components and fermentation conditions were evaluated in this work.ResultsThe by-products analysis showed that there existed reversible transformation between acetoin and 2,3-butanediol that was strictly responsible for acetoin production in B. subtilis SF4-3. The carbon sources, nitrogen sources and agitation speed were determined to play crucial role in the acetoin production. The optimal media (glucose·H2O 150 g/L, yeast extract 10 g/L, corn steep dry 5 g/L, urea 2 g/L, K2HPO4 0.5 g/L, MgSO4 0.5 g/L) were obtained. Furthermore, the low agitation speed of 300 r/min was found to be beneficial to the reversible transformation of 2,3-butanediol for acetoin production in B. subtilis SF4-3. Eventually, 48.9 g/L of acetoin and 5.5 g/L of 2,3-butanediol were obtained in a 5-L fermenter, and the specific production of acetoin was 39.12% (g/g), which accounted for 79.90% of the theoretical conversion.ConclusionsThe results indicated acetoin production of B. subtilis SF4-3 was closely related to the medium components and dissolved oxygen concentrations. It also provided a method for acetoin production via the reversible transformation of acetoin and 2,3-butanediol.  相似文献   

5.
Background1,3-Propanodiol (1,3-PD), is used in the production of polytrimethylene terephthalate (PTT), an aromatic polyester that exhibits high elastic recoveries. It is also employed as a supplement with low solidification properties, a solvent and a lubricant in the formof propylene glycol. 1,3-PD is effectively synthesized by a microbiological way from crude glycerol. The main problem of this technology is using a high concentration of glycerol, which is a limiting factor for bacteria cells growth (especially in batch fermentation).ResultsIn this work, the influence of different glycerol concentration in batch fermentation on Clostridium butyricum DSP1 metabolism was investigated. The biomass was concentrated for two times with the use of membrane module (in case of increasing kinetic parameters). Increased optical density of bacteria cells six times increased the productivity of 1,3-PD in cultivation with 20 g/L of glycerol at the beginning of the process, and more than two times in cultivation with 60–80 g/L. Also the possibility of complete attenuation of 140 g/L of crude glycerol in the batch fermentation was investigated. During the cultivation, changes of protein profiles were analyzed. The most significant changes were observed in the cultivation in the medium supplemented with 80 g/L of glycerol. They related mainly to the DNA protein reconstructive systems, protective proteins (HSP), and also the enzymatic catalysts connected with glycerol metabolic pathway.ConclusionsThe application of filtration module in batch fermentation of crude glycerol by C. butyricum DSP1 significantly increased the productivity of the process.  相似文献   

6.
BackgroundThe development of a potential single culture that can co-produce hydrogen and ethanol is beneficial for industrial application. Strain improvement via molecular approach was proposed on hydrogen and ethanol co-producing bacterium, Escherichia coli SS1. Thus, the effect of additional copy of native hydrogenase gene hybC on hydrogen and ethanol co-production by E. coli SS1 was investigated.ResultsBoth E. coli SS1 and the recombinant hybC were subjected to fermentation using 10 g/L of glycerol at initial pH 7.5. Recombinant hybC had about 2-fold higher cell growth, 5.2-fold higher glycerol consumption rate and 3-fold higher ethanol productivity in comparison to wild-type SS1. Nevertheless, wild-type SS1 reported hydrogen yield of 0.57 mol/mol glycerol and ethanol yield of 0.88 mol/mol glycerol, which were 4- and 1.4-fold higher in comparison to recombinant hybC. Glucose fermentation was also conducted for comparison study. The performance of wild-type SS1 and recombinant hybC showed relatively similar results during glucose fermentation. Additional copy of hybC gene could manipulate the glycerol metabolic pathway of E. coli SS1 under slightly alkaline condition.ConclusionsHybC could improve glycerol consumption rate and ethanol productivity of E. coli despite lower hydrogen and ethanol yields. Higher glycerol consumption rate of recombinant hybC could be an advantage for bioconversion of glycerol into biofuels. This study could serve as a useful guidance for dissecting the role of hydrogenase in glycerol metabolism and future development of effective strain for biofuels production.  相似文献   

7.
BackgroundThe production of biofuels from renewable energy sources is one of the most important issues in industrial biotechnology today. The process is known to generate various by-products, for example crude glycerol, which is obtained in the making of biodiesel from rapeseed oil. Crude glycerol may be utilized in many ways, including microbial conversion to 1,3-propanediol (1,3-PD), a raw material for the synthesis of polyesters and polyurethanes.ResultsThe paper presents results of a study on the synthesis of 1,3-propanediol from crude glycerol by a repeated batch method with the use of Clostridium butyricum DSP1. Three cycles of fermentation medium replacement were carried out. The final concentration of 1,3-PD was 62 g/L and the maximum productivity, obtained during the second cycle, reached 1.68 g/L/h. Additionally, experiments conducted in parallel to the above involved using the entire quantity of the culture broth removed from the bioreactor to inoculate successive portions of fermentation media containing crude glycerol at concentrations of 80 g/L and 100 g/L. Under those conditions, the maximum 1,3-PD concentrations were 43.2 g/L and 54.2 g/L.ConclusionsThe experiments proved that by using a portion of metabolically active biomass as inoculum for another fermentation formula it is possible to eliminate the stage of inoculum growth and thereby reduce the length of the whole operation. Additionally, that strategy avoids the phase of microbial adaptation to a different source of carbon such as crude glycerol, which is more difficult to utilize, thus improving the kinetic parameters of 1,3-PD production.  相似文献   

8.
BackgroundAn effective single culture with high glycerol consumption and hydrogen and ethanol coproduction yield is still in demand. A locally isolated glycerol-consuming Escherichia coli SS1 was found to produce lower hydrogen levels under optimized ethanol production conditions. Molecular approach was proposed to improve the hydrogen yield of E. coli SS1 while maintaining the ethanol yield, particularly in acidic conditions. Therefore, the effect of an additional copy of the native hydrogenase gene hycE and recombinant clostridial hydrogenase gene hydA on hydrogen production by E. coli SS1 at low pH was investigated.ResultsRecombinant E. coli with an additional copy of hycE or clostridial hydA was used for fermentation using 10 g/L (108.7 mmol/L) of glycerol with an initial pH of 5.8. The recombinant E. coli with hycE and recombinant E. coli with hydA showed 41% and 20% higher hydrogen yield than wild-type SS1 (0.46 ± 0.01 mol/mol glycerol), respectively. The ethanol yield of recombinant E. coli with hycE (0.50 ± 0.02 mol/mol glycerol) was approximately 30% lower than that of wild-type SS1, whereas the ethanol yield of recombinant E. coli with hydA (0.68 ± 0.09 mol/mol glycerol) was comparable to that of wild-type SS1.ConclusionsInsertion of either hycE or hydA can improve the hydrogen yield with an initial pH of 5.8. The recombinant E. coli with hydA could retain ethanol yield despite high hydrogen production, suggesting that clostridial hydA has an advantage over the hycE gene in hydrogen and ethanol coproduction under acidic conditions. This study could serve as a useful guidance for the future development of an effective strain coproducing hydrogen and ethanol.  相似文献   

9.
BackgroundGABA (γ-aminobutyric acid) is a four-carbon nonprotein amino acid that has hypotensive, diuretic, and tranquilizing properties. Glutamate decarboxylase (GAD) is the key enzyme to generate GABA. A simple and economical method of preparing and immobilizing GAD would be helpful for GABA production. In this study, the GAD from Lactobacillus fermentum YS2 was expressed under the control of a stress-inducible promoter and was purified and immobilized in a fusion form, and its reusability was investigated.ResultsThe fusion protein CBM-GAD was expressed in Escherichia coli DH5α carrying pCROCB-gadB, which contained promoter PrpoS, cbm3 (family 3 carbohydrate-binding module from Clostridium thermocellum) coding sequence, the gadB gene from L. fermentum YS2 coding for GAD, and the T7 terminator. After a one-step purification of CBM-GAD using regenerated amorphous cellulose (RAC) as an adsorbent, SDS-PAGE analysis revealed a clear band of 71 kDa; the specific activity of the purified fusion protein CBM-GAD reached 83.6 ± 0.7 U·mg-1. After adsorption onto RAC, the immobilized GAD with CBM3 tag was repeatedly used for GABA synthesis. The protein-binding capacity of RAC was 174 ± 8 mg·g-1. The immobilized CBM-GAD could repeatedly catalyze GABA synthesis, and 8% of the initial activities was retained after 10 uses. We tested the conversion of monosodium glutamate to GABA by the immobilized enzyme; the yield reached 5.15 g/L and the productivity reached 3.09 g/L·h.ConclusionsRAC could be used as an adsorbent in one-step purification and immobilization of CBM-GAD, and the immobilized enzyme could be repeatedly used to catalyze the conversion of glutamate to GABA.  相似文献   

10.
BackgroundCatalase (CAT) is an important enzyme that degrades H2O2 into H2O and O2. To obtain an efficient catalase, in this study, a new strain of high catalase-producing Serratia marcescens, named FZSF01, was screened and its catalase was purified and characterized.ResultsAfter optimization of fermentation conditions, the yield of catalase produced by this strain was as high as 51,468 U/ml. This catalase was further purified using two steps: DEAE-fast flow and Sephedex-G150. The purified catalase showed a specific activity of 197,575 U/mg with a molecular mass of 58 kDa. This catalase exhibited high activity at 20–70°C and pH 5.0–11.0. Km of the catalase was approximately 68 mM, and Vmax was 1886.8 mol/min mg. This catalase was further identified by LC–MS/MS, and the encoding gene was cloned and expressed in Escherichia coli BL21 (DE3) with a production of 17,267 ± 2037 U/ml.ConclusionsTo our knowledge, these results represent one of the highest fermentation levels reported among current catalase-producing strains. This FZSF01 catalase may be suitable for several industrial applications that comprise exposure to alkaline conditions and under a wide range of temperatures.  相似文献   

11.
BackgroundMucor indicus is a dimorphic fungus used in the production of ethanol, oil, protein, and glucosamine. It can ferment different pentoses and hexoses; however, the yields of products highly depend on the nutrients and cultivation conditions. In this study, the effects of different morphologic forms, cultivation time and temperature, presence or absence of oxygen, carbon sources, and concentration of nitrogen source on the products of M. indicus were investigated.ResultsThe fungus with all morphologies produced high yields of ethanol, in the range of 0.32–0.43 g/g, on glucose. However, the fungus with filamentous morphology produced higher amounts of oil, protein, phosphate, and glucosamine together with ethanol, compared with other morphologies. A higher amount of oil (0.145 g/g biomass) was produced at 28°C, while the best temperature for protein and glucosamine production was 32 and 37°C, respectively. Although ethanol was produced at a higher yield (0.44 g/g) under anaerobic conditions compared with aerobic conditions (yield of 0.41 g/g), aerobic cultivation resulted in higher yields of protein (0.51 g/g biomass), glucosamine (0.16 g/g alkali insoluble material, AIM), and phosphate (0.11 g/g AIM).ConclusionsIt is not possible to have the maximum amounts of the products simultaneously. The fermentation conditions and composition of culture media determine the product yields. Carbon source type and the addition of nitrogen source are among the most influencing factors on the product yields. Moreover, all measured products were made with higher yields in cultivation on glucose, except glucosamine, which was produced with higher yields on xylose.  相似文献   

12.
BackgroundRhodotorula glutinis is capable of synthesizing numerous valuable metabolites with extensive potential industrial usage. This paper reports the effect of initial culture medium pH on growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.ResultsThe highest biomass yield was obtained in media with pH 4.0–7.0, and the value after 72 h was 17.2–19.4 gd.w./L. An initial pH of the medium in the range of 4.0–7.0 has no significant effect on the protein (38.5–41.3 g/100 gd.w.), lipid (10.2–12.7 g/100 gd.w.), or carotenoid (191.7–202.9 μg/gd.w.) content in the biomass or on the profile of synthesized fatty acids and carotenoids. The whole pool of fatty acids was dominated by oleic (48.1–53.4%), linoleic (21.4–25.1%), and palmitic acids (13.0–15.8%). In these conditions, the yeast mainly synthesized torulene (43.5–47.7%) and β-carotene (34.7–38.6%), whereas the contribution of torularhodin was only 12.1–16.8%. Cultivation in medium with initial pH 3.0 resulted in a reduction in growth (13.0 gd.w./L) and total carotenoid (115.8 μg/gd.w.), linoleic acid (11.5%), and torularhodin (4.5%) biosynthesis.ConclusionThe different values of initial pH of the culture medium with glycerol and deproteinized potato wastewater had a significant effect on the growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.  相似文献   

13.
BackgroundPoly(dl-lactic acid), or PDLLA, is a biodegradable polymer that can be hydrolyzed by various types of enzymes. The protease produced by Actinomadura keratinilytica strain T16-1 was previously reported to have PDLLA depolymerase activity. However, few studies have reported on PDLLA-degrading enzyme production by bacteria. Therefore, the aims of this study were to determine a suitable immobilization material for PDLLA-degrading enzyme production and optimize PDLLA-degrading enzyme production by using immobilized A. keratinilytica strain T16-1 under various fermentation process conditions in a stirrer fermenter.ResultsAmong the tested immobilization materials, a scrub pad was the best immobilizer, giving an enzyme activity of 30.03 U/mL in a shake-flask scale. The maximum enzyme activity was obtained at aeration 0.25 vvm, agitation 170 rpm, 45°C, and 48 h of cultivation time. Under these conditions, a PDLLA-degrading enzyme production of 766.33 U/mL with 15.97 U/mL·h productivity was observed using batch fermentation in a 5-L stirrer fermenter. Increased enzyme activity and productivity were observed in repeated-batch (942.67 U/mL and 19.64 U/mL·h) and continuous fermentation (796.43 U/mL and 16.58 U/mL·h) at a dilution rate of 0.013/h. Scaled-up production of the enzyme in a 10-L stirrer bioreactor using the optimized conditions showed a maximum enzyme activity of 578.67 U/mL and a productivity of 12.06 U/mL·h.ConclusionsThis research successfully scaled-up the enzyme production to 5 and 10 L in a stirrer fermenter and is helpful for many applications of poly(lactic acid).  相似文献   

14.
BackgroundSurfactants are one of the most important raw materials used in various industrial fields as emulsifiers, corrosion inhibitors, foaming agents, detergent products, and so on. However, commercial surfactant production is costly, and its demand is steadily increasing. This study aimed to evaluate the performance of typical strains of Bacillus sp. to produce biosurfactants through fermentation. It also included the investigation of the effect of initial glucose concentration and the carbon to nitrogen ratio.ResultsThe biosurfactant yield was in the range of 1–2.46 g/L at initial glucose concentrations of 10–70 g/L. The optimum fermentation condition was achieved at a carbon to nitrogen ratio of 12.4, with a decrease in surface tension of up to 27 mN/m.ConclusionsFor further development and industrial applications, the modified Gompertz equation is proposed to predict the cell mass and biosurfactant production as a goodness of fit was obtained with this model. The modified Gompertz equation was also extended to enable the excellent prediction of the surface tension.  相似文献   

15.
BackgroundBiohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated.ResultsThe methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15–20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P < 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P < 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield.ConclusionPreventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.  相似文献   

16.
BackgroundThe acidic subunit of amarantin (AAC)—the predominant amaranth seed storage protein—has functional potential and its third variable region (VR) has been modified with antihypertensive peptides to improve this potential. Here, we modified the C-terminal in the fourth VR of AAC by inserting four VY antihypertensive peptides. This modified protein (AACM.4) was expressed in Escherichia coli. In addition, we also recombinantly expressed other derivatives of the amarantin protein. These include: unmodified amarantin acidic subunit (AAC); amarantin acidic subunit modified at the third VR with four VY peptides (AACM.3); and amarantin acidic subunit doubly modified, in the third VR with four VY peptides and in the fourth VR with the RIPP peptide (AACM.3.4).ResultsE. coli BL21-CodonPlus (DE3)-RIL was the most favorable strain for the expression of proteins. After 6 h of induction, it showed the best recombinant protein titer. The AAC and AACM.4 were obtained at higher titers (0.56 g/L) while proteins modified in the third VR showed lower titers: 0.44 g/L and 0.33 g/L for AACM.3 and AACM.3.4, respectively. As these AAC variants were mostly expressed in an insoluble form, we applied a refolding protocol. This made it possible to obtain all proteins in soluble form. Modification of the VR 4 improves the thermal stability of amarantin acidic subunit; AAC manifested melting temperature (Tm) at 34°C and AACM.4 at 37.2°C. The AACM.3 and AACM.3.4 did not show transition curves.ConclusionsModifications to the third VR affect the thermal stability of amarantin acidic subunit.  相似文献   

17.
BackgroundThe selection of new yeast strains could lead to improvements in bioethanol production. Here, we have studied the fermentative capacity of different auxotrophic mutants of Saccharomyces cerevisiae, which are routinely used as hosts for the production of heterologous proteins. It has recently been found that these strains exhibit physiological alterations and peculiar sensitivities with respect to the parental prototrophic strains from which they derive. In this work the performance of auxotrophic S. cerevisiae CEN.PK strains was compared to the corresponding prototrophic strain, to S. cerevisiae T5bV, a strain isolated from grape must and to another auxotrophic strain, S. cerevisiae BY4741.ResultsThe results indicate that the fermentative capacity of strains grown in 2% glucose was similar in all the strains tested. However, in 15% initial glucose, the auxotrophic strains exhibited a more than doubled ethanol yield on biomass (10 g g- 1dw) compared to the prototrophic strains (less than 5 g g- 1dw). Other tests have also evidenced that in medium depletion conditions, ethanol production continues after growth arrest.ConclusionsThe results highlight the capacity of auxotrophic yeast strains to produce ethanol per mass unit, in a higher amount with respect to the prototrophic ones. This leads to potential applications for auxotrophic strains of S. cerevisiae in the production of ethanol in both homogeneous and heterogeneous phases (immobilized systems). The higher ethanol yield on biomass would be advantageous in immobilized cell systems, as a reduced yeast biomass could greatly reduce the mass transfer limitations through the immobilization matrix.  相似文献   

18.
BackgroundEndoglucanase plays a major role in initiating cellulose hydrolysis. Various wild-type strains were searched to produce this enzyme, but mostly low extracellular enzyme activities were obtained. To improve extracellular enzyme production for potential industrial applications, the endoglucanase gene of Bacillus subtilis M015, isolated from Thai higher termite, was expressed in a periplasmic-leaky Escherichia coli. Then, the crude recombinant endoglucanase (EglS) along with a commercial cellulase (Cel) was used for hydrolyzing celluloses and microbial hydrolysis using whole bacterial cells.ResultsE. coli Glu5 expressing endoglucanase at high levels was successfully constructed. It produced EglS (55 kDa) with extracellular activity of 18.56 U/mg total protein at optimal hydrolytic conditions (pH 4.8 and 50°C). EglS was highly stable (over 80% activity retained) at 40–50°C after 100 h. The addition of EglS significantly improved the initial sugar production rates of Cel on the hydrolysis of carboxymethyl cellulose (CMC), microcrystalline cellulose, and corncob about 5.2-, 1.7-, and 4.0-folds, respectively, compared to those with Cel alone. E. coli Glu5 could secrete EglS with high activity in the presence of glucose (1% w/v) and Tween 80 (5% w/v) with low glucose consumption. Microbial hydrolysis of CMC using E. coli Glu5 yielded 26 mg reducing sugar/g CMC at pH 7.0 and 37°C after 48 h.ConclusionsThe recombinant endoglucanase activity improved by 17 times compared with that of the native strain and could greatly enhance the enzymatic hydrolysis of all studied celluloses when combined with a commercial cellulase.  相似文献   

19.
BackgroundBiomineralization is a significant process performed by living organisms in which minerals are produced through the hardening of biological tissues. Herein, we focus on calcium carbonate precipitation, as part of biomineralization, to be used in applications for environmental protection, material technology, and other fields. A strain GM-1, Microbacterium sp. GM-1, isolated from active sludge, was investigated for its ability to produce urease and induce calcium carbonate precipitation in a metabolic process.ResultsIt was discovered that Microbacterium sp. GM-1 resisted high concentrations of urea up to 60 g/L. In order to optimize the calcification process of Microbacterium sp. GM-1, the concentrations of Ni2 + and urea, pH value, and culture time were analyzed through orthogonal tests. The favored calcite precipitation culture conditions were as follows: the concentration of Ni2 + and urea were 50 μM and 60 g/L, respectively, pH of 10, and culture time of 96 h. Using X-ray diffraction analysis, the calcium carbonate polymorphs produced by Microbacterium sp. GM-1 were proven to be mainly calcite.ConclusionsThe results of this research provide evidence that Microbacterium sp. GM-1 can biologically induce calcification and suggest that strain GM-1 may play a potential role in the synthesis of new biominerals and in bioremediation or biorecovery.  相似文献   

20.
BackgroundLysozyme plays a crucial role in innate immunity with its well-recognized bacteriolytic activity. In this study, the influence of expression parameters (inoculation volume, culture volume, growth time, induction temperature and time, initial pH and methanol concentration) on human lysozyme (HLZ) production in recombinant P. pastoris SMD1168 was investigated through Plackett–Burman (PB) design and response surface methodology (RSM).ResultsIt was revealed that induction temperature, induction time and culture volume had significant influence (P < 0.01) on HLZ expression level, which were elected for further optimization with three-dimensional response surface designs for enhanced HLZ production. The highest lysozyme activity reached 3301 U/mL under optimized conditions (at 23.5°C for 90 h with culture volume of 48 mL) in shake flask, which increased 2.2 fold compared with that achieved with the standard protocol (Invitrogen). When high-cell-density fermentation of the recombinant Pichia pastoris was performed in a 15 L fermenter under optimized conditions, the extracellular lysozyme activity reached 47,680 U/mL. SDS-PAGE analysis of the product demonstrated that HLZ was produced as a single major protein with a molecular weight of approximately 14.7 kDa, consistent with its expected size.ConclusionsThe results indicated that the optimized culture conditions using PB design and RSM significantly enhanced the expression level of HLZ, and the Pichia expression system for HLZ production was successful and industrially promising.  相似文献   

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