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1.
Takeshi Omasa Masaru Ishimoto Ken-ichi Higashiyama Suteaki Shioya Ken-ichi Suga 《Cytotechnology》1992,8(1):75-84
The concentration effects of certain amino acids (Asp, Ile, Leu, Lys, Met, Val, Phe and Gln which were highly consumed during
cultivation), and glucose on cell growth and antibody productivity were investigated using dish culture. From these experiments,
it was found that only glutamine enrichment enhanced the specific antibody production rate. The other amino acids described
above did not affect either the specific growth rate or specific antibody production rate. Thus we investigated the quantitative
effects of glutamine concentration in the range of 0.4∼33.3 mmol·1−1 on kinetic parameters in fed-batch culture which kept both glucose and glutamine concentration constant. As a result the
specific growth rate decreased with increase in glutamine concentration in the range larger than 20 mmol·1−1. The specific antibody production rate had a maximum value at about 25 mmol·1−1 glutamine concentration. 相似文献
2.
Yaw-Nan Chang Jen-Chang Huang Chih-Chen Lee Ing-Lung Shih Yew-Min Tzeng 《Enzyme and microbial technology》2002,30(7):314-894
Response surface methodology (RSM) was employed to study the effect of culture medium on the production of lovastatin in mixed solid-liquid state (or submerged) cultures by Monascus ruber. The maximal lovastatin yield (131 mg/L, average of three repeats) appeared at the region where the respective concentrations of rice powder, peptone, glycerin, and glucose were around 34.4 g/L, 10.8 g/L, 26.4 ml/L, and 129.2 g/L, respectively. The optimized medium resulted in a significant increase of lovastatin yield, as compared with that obtained by the fermentation of many other M. ruber species. 相似文献
3.
Francis Mairet Pierre Villon Michèle Boitel‐Conti Khalil Shakourzadeh 《Biotechnology progress》2010,26(3):847-856
This article proposes a feeding strategy based on a kinetic model to enhance hairy roots growth. A new approach for modeling hairy root growth is used, considering that there is no nutrient limitation thanks to an appropriate feeding, and the intracellular pools are supposed to be always saturated. Thus, the model describes the specific growth rate from extracellular concentration of the major nutrients and nutrient uptakes depend on biomass growth. An optimized feeding strategy was determined thanks to the model to maintain the major nutrient levels at their optimum assuming optimal initial concentrations. The optimal feed rate is computed in open loop using kinetic model prediction or in closed loop using conductivity measurements to estimate biomass growth. Datura innoxia was chosen as the model culture system. Shake flask cultures were used to calibrate the model. Finally, cultures in bioreactor were performed to validate the model and the control laws. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 相似文献
4.
Methanobacterium thermoautotrophicum, strain Hveragerdi, has been cultivated in a completely defined mineral salts medium, under strictly anaerobic conditions with CO2 and H2 as sole carbon and energy source, respectively. During optimization of the medium an iron limitation was identified that could not be overcome by the simple addition of iron—or iron complexed with nitrilotriacetate (NTA)—to the medium, due to the formation of insoluble FeS complexes. In order to define a medium optimization strategy, and to avoid laborious empirical optimization procedures, a theoretical model has been developed in order to describe the solubility of iron and other mineral species in the medium as a function of the concentration of sulfur and NTA. This model may be applied for the optimization of any medium component. With this information sulfide has been replaced by a combination of cysteine and thiosulphate in conjunction with a non-toxic reducing agent (titanium (III) citrate). Using this defined medium precipitation was avoided and an iron limitation was overcome resulting in a 5-fold improvement of the final biomass concentration from 2–3 g l−1 to 11.2 g l−1 together with a 2-fold increase (from 45 to almost 100%) in the conversion of CO2 and H2 to CH4, even at gas flow rates as high as 6 l min−1. 相似文献
5.
Olfa Tabbene Imen Ben Slimene Kais Djebali Maria‐Luisa Mangoni Maria‐Camino Urdaci Ferid Limam 《Biotechnology progress》2009,25(5):1267-1274
An antimicrobial activity produced by Bacillus subtilis B38 was found to be effective against several bacteria, including pathogenic and spoilage microorganisms such as, Listeria monocytogenes, Salmonella enteridis, and clinical isolates of methicillin‐resistant Staphylococcus species. Nutrients such as carbon, nitrogen sources, and inorganic salts enhanced the production level of the antibacterial activity by B. subtilis B38. A first screening step showed that lactose, ammonium succinate, and manganese most influenced both cell growth and antibacterial activity production. These three factors varied at two levels in eight experiments using full factorial design. Results indicated that maximum cell growth (OD = 10.2) and maximum production of antibacterial activity (360 AU/mL) were obtained in a modified medium containing 1.5% (w/v) lactose, 0.15% (w/v) ammonium succinate, and 0.3 mg/L manganese. Depending on the indicator strain used, the antibacterial activity was 2‐ to 4‐fold higher in the modified culture medium than in TSB medium under the same conditions. Thin layer chromatography‐bioautography assay showed the presence of three active spots with Rf values of 0.47, 0.7, and 0.82 in TSB medium. However, the inhibition zone of two spots (Rf values of 0.7 and 0.82) was slightly larger in the modified medium. Moreover, a large zone of inhibition with an Rf value of 0.3, was observed in this modified medium, instead of the spot having an Rf value of 0.47. These results suggest that the nutrients act as environmental factors, quantitatively and qualitatively affecting the production of antibacterial compounds by B. subtilis B38. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 相似文献
6.
Use of response surface methodology to optimize culture medium for production of lipase with Candida sp. 99-125 总被引:1,自引:0,他引:1
Response surface methodology (RSM) was employed to optimize culture medium for production of lipase with Candida sp. 99-125. In the first step, a Plackett–Burmen design was used to evaluate the effects of different components in the culture medium. Soybean oil, soybean powder and K2HPO4 have significant influences on the lipase production. The concentrations of three factors were optimized subsequently using central composite designs and response surface analysis. The optimized condition allowed the production of lipase to be increased from 5000 to 6230 IU/ml in shake flask system. The lipase fermentation in 5 l fermenter reached 9600 IU/ml. 相似文献
7.
Moran EB McGowan ST McGuire JM Frankland JE Oyebade IA Waller W Archer LC Morris LO Pandya J Nathan SR Smith L Cadette ML Michalowski JT 《Biotechnology and bioengineering》2000,69(3):242-255
A systematic approach to the validation of control ranges of control parameters for a cell culture process producing a monoclonal antibody is described. Specifically, the structure and functional activity of a monoclonal IgG1 antibody produced at the outer limits of numerical ranges of fed-batch culture control parameters such as pH and temperature were examined, with the aim of providing assurance that antibody produced under varying culture conditions was of consistent quality based on a carefully defined set of specifications. An experimental design was created using a half-fractional factorial design for fed-batch culture incorporating half of the thirty two possible combinations of five selected control parameters at high and low levels. Statistical analysis of all data gathered from the study allowed an assessment of the effects of the process control parameters at either high or low outer limits on fed-batch culture response variables such as growth rate and specific antibody productivity. Measured values for the responses of growth rate and specific antibody productivity throughout this study ranged from 0.22-0.44 d(-1) and 6.4-32 microg monoclonal antibody/10(6) cells/d respectively. Analytical characterisation of monoclonal antibody purified from each fed-batch culture considered the purity, structure and biological activity of the glycoprotein. All antibody preparations were identical to each other and to the current antibody reference standard or control. Glycosylation analysis of certain samples from the study demonstrated that the distribution of glycoforms of the antibody was not affected by the varying process control conditions of the fed-batch cultures. 相似文献
8.
Jianguang Luo Jun Liu Chunling Ke Deliang Qiao Hong Ye Yi Sun Xiaoxiong Zeng 《Carbohydrate polymers》2009,78(3):409-415
Optimization of medium composition for the production of exopolysaccharides (EPS) from Phellinus baumii Pilát in submerged culture and the immuno-stimulating activity of EPS were carried out. Firstly, the medium components having significant effect on EPS production were screened out to be glucose, yeast extract and diammonium oxalate monohydrate by using a 2(7−3) fractional factorial design. Secondly, the concentrations of the three factors were optimized using central composite design in response surface methodology. As results, a quadratic model was found to fit for EPS production, and the optimal medium composition was determined as following (g/l): 34.12 glucose, 4 peptone, 5.01 yeast extract, 0.88 diammonium oxalate monohydrate, 0.75 MgSO4 and 1 KH2PO4 and 0.0075 thiamine (VB1). A yield of 2.363 ± 0.04 g/l for EPS was observed in verification experiment. Finally, EPS from P. baumii Pilát was found to have direct immuno-stimulating activity in vitro on splenocyte proliferative response and acid phosphatase activity in peritoneal macrophages in a dose-dependent manner. 相似文献
9.
High-cell-density production of recombinant growth hormone of Lateolabrax japonicus (rljGH) expressed intracellularly in Pichia pastoris was investigated. In the regular strategy of induction at a cell density of 160 g l−1, short duration of intracellular rljGH accumulation (17 h) resulted in a low final cell density of 226 g l−1. Thus, a novel strategy of induction at a cell density of 320 g l−1 was investigated. In this strategy, the preinduction glycerol-feeding scheme had a significant effect on the post-induction
production. Constant glycerol feeding led to a decrease of the specific rljGH production and specific production rate because
of low preinduction specific growth rate. This decrease was avoided by exponential glycerol feeding to maintain a preinduction
specific growth rate of 0.16 h−1. The results from exponential glycerol feeding indicated that the rljGH production depended on the preinduction specific
growth rate. Moreover, mixed feeding of methanol and glycerol during induction improved the specific production rate to 0.07 mg
g−1 h−1 from 0.043 mg g−1 h−1. Consequently, both high cell density (428 g l−1) and high rljGH production could be achieved by the novel strategy: growing the cells at the specific growth rate of 0.16 h−1 to the cell density of 320 g l−1 and inducing the expression by mixed feeding. 相似文献
10.
Xiao-Ling Tang Zhi-Chao Wang Jian Yang Yu-Guo Zheng 《Preparative biochemistry & biotechnology》2013,43(2):117-126
AbstractTyrosine phenol lyase (TPL) is a robust biocatalyst for the production of L-dihydroxyphenylalanine (L-DOPA). The improvement of TPL production is conducive to the industrial potential. In this study, the optimization of culture medium of recombinant Escherichia coli harboring TPL from Fusobacterium nucleatum (Fn-TPL) was carried out. Sucrose and combination of yeast extract and peptone were selected as carbon and nitrogen source, respectively. Their optimal concentrations were determined by Box-Behnken design and the synergistic effect between yeast extract and peptone was found to be significant, with p-value < 0.05. The DO-STAT fed-batch fermentation under optimized culture condition was established and the oxygen level was fixed at 20%. Both the biomass and Fn-TPL activity were significantly increased, which were 35.6 g dcw/L and 12292 U/L, respectively. The results obtained significantly promote the industrial production of L-DOPA production. 相似文献
11.
Effect of medium composition and culture conditions on agarase production by Agarivorans albus YKW-34 was investigated in shake flasks. The most suitable carbon source, nitrogen source, and culture temperature were agar, yeast extract, and 25 °C, respectively, for agarase production by one-factor-at-a-time design. The nutritional components of the medium and culture conditions were analyzed by Plackett–Burman design. Among the nine factors studied, agar, yeast extract, and initial pH had significant effects on agarase production (p < 0.05). The optimum levels of these key variables were further determined using a central composite design. The highest agarase production was obtained in the medium consisting of 0.23% agar and 0.27% yeast extract at initial pH 7.81. The whole optimization strategy enhanced the agarase production from 0.23 U/ml to 0.87 U/ml. The economic medium composition and culture condition as well as the dominant occupation of agarase with high activity in culture fluid enlighten the potential application of A. albus YKW-34 for the production of agarase. 相似文献
12.
Abstract Oleandrin has been identified as the most potent antitumor ingredient of the Mediterranean herb Nerium oleander L. A strategy for optimization of medium compositions and conditions was developed for enhanced oleandrin production in suspension cultures from leaf-origin explants of Nerium oleander. The cell suspension cultures were grown in various modifications of MS medium as a basal medium. The effects of different natural extracts, plant growth substances, carbon and nitrogen sources and phosphate on the growth and oleandrin accumulation were investigated as well as effect of light, pH, shaking speed and substrate feeding. The highest oleandrin yield was obtained when the nitrogen concentration was lowered to two-thirds and the phosphate concentration increased by two-thirds of that specified in the MS medium in the presence of 3% sucrose, coconut milk, indolebutyric acid and benzyladenine in concentrations of 1 and 2 mg l?1, respectively. Lower pH and faster shaking speed favored oleandrin accumulation. Chemical feeding of progesterone and cholesterol boosted the oleandrin concentration to higher levels reaching 8.23±0.05 mg g?1 dry weight. This was about 10-fold higher than that detected in field-grown plants using the same extraction and analytic conditions, and about 24-fold higher than that determined in control cultures with regular MS medium and without precursor feeding. 相似文献
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14.
A new parameter could be introduced to facilitate the optimization of media used for cultivation of stock cultures on agar slants. This parameter reduces the amount of data generated in optimization experiments to one single value (hs-value) for each medium composition. The hs-value (high and stable product formation) allows an assessment of any medium formulation with regard to reproducibility and product formation, demonstrated for the production process of the antibiotic gallidermin by Staphylococcus gallinarum TÜ 3928. © Rapid Science Ltd. 1998 相似文献
15.
Meng-Hao Cai Xiang-Shan Zhou Xue-Qian Sun Ke-Jing Tao Yuan-Xing Zhang 《Journal of industrial microbiology & biotechnology》2009,36(3):381-389
Statistical methodologies were employed to optimize submerged culture medium for the production of a novel antineoplastic
compound aspergiolide A by a marine-derived fungus Aspergillus glaucus HB1-19 for the first time. Orthogonal design was preformed to determine the initial composition. Then Plackett–Burman design
was applied to evaluate the influence of related nutrients, and yeast extract paste, soybean powder and sodium glutamate were
confirmed as critical factors in the medium. Response surface methodology (RSM) was finally taken as an effective approach
to optimize the combination of the obtained three significant factors. The predicted maximal aspergiolide A production of
62.4 mg/L appeared at the region where the concentrations of sodium glutamate, soybean powder, and yeast extract paste were
2, 1, and 1.07 g/L, respectively. Under the proposed optimized conditions, the experimental aspergiolide A production reached
71.2 mg/L. The correlation between predicted value and measured value of these experiments proved the validity of the response
model. After optimization, aspergiolide A production increased 4.22 times compared to that of the original medium. Elemental
analysis was finally taken into consideration, and carbon–nitrogen ratio in the medium increased from 20.1:1 to 86.6:1. This
great difference was inferred as the most important reason for production enhancement by metabolic pathway analysis. 相似文献
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17.
Hyper-expression of a secretory exoglucanase, Exg, encoded by the cex gene of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of recombinant Escherichia coli (Z.B. Fu, K.L. Ng, T.L. Lam, W.K.R. Wong, Cell death caused by hyper-expression of a secretory exoglucanase in Esherichia coli, Protein Expr. Purif. 42 (2005) 67-77). We propose here that the cell lysate ratio (Pre/Mat RQ) of the unprocessed precursor Exg protein (Pre-Exg) and its processed mature product (Mat-Exg) reflects the capacity of E. coli to secrete Exg. A Pre/Mat RQ of 20/80, designated the "Critical Value," was an important threshold measurement. A rise in the Pre/Mat RQ triggered a mass killing effect. The use of various secretion signal peptides did not improve the viability of cells expressing high levels of Pre-Exg under strong tac promoter control. However, use of the weaker vegG promoter in conjunction with a change in start codon of the spa leader sequence from ATG to TTG in a pM1vegGcexL plasmid construct resulted in a high level (0.9 U ml(-1)) of excreted Exg in shake-flask cultures. This was 50% higher than the best result obtained from plasmid construct lacUV5par8cex, using the lacUV5 promoter and the ompA leader sequence. Variations in the excreted Exg activities were attributable to differences in the Pre/Mat RQ values of the induced cultures harboring pM1vegGcexL and lacUV5par8cex. These values were 18/82 and 10/90, respectively. Employing fed-batch cultivation in two-liter fermentors, an induced JM101(pM1vegGcexL) culture yielded 4.5 U ml(-1) of excreted Exg, which was over six fold greater that previously reported. Our results illustrate the successful application of the Pre/Mat RQ ratio as a guide to the attainment of a maximum level of secreted/excreted Exg. 相似文献
18.
The optimum experimental conditions (sample size and mobile phase composition) are calculated for maximum production rate of either one of two enantiomers contained in feeds of different compositions (1/1, 1/10, and 10/1). The products are obtained at 99% purity. The calculations use the equilibrium-dispersive model of chromatography and the equilibrium isotherms determined experimentally from the rear, diffuse boundary of overloaded elution profiles. The production rate measured experimentally under the optimum conditions calculated agree with 4% of the calculated values. There is an optimum value for the retention factor which is higher than predicted by a model assuming constant separation factor, because both separation factor and retention decrease with increasing organic solvent concentration in the mobile phase. (c) 1992 John Wiley & Sons, Inc. 相似文献
19.
Key message
A method of combination of ingrowth core and scanner provides better estimation of fine root production.Abstract
Estimating fine root (diameter ≤2 mm) production has been difficult. The best method remains unknown because each method has potential biases associated with the estimation methods and field techniques applied, leading to over- or underestimation. In this study, a new method combining ingrowth core and scanner named as “combined method” was described, and it was applied to a Quercus serrata plantation in Tsukuba, Japan. Comparing accuracy of estimation of fine root production by decision matrix and continuous inflow method using ingrowth core with that by “combined method” was conducted. Closed-box protected scanners and ingrowth cores were buried in a field on December 6, 2012, and fine root images were nondestructively scanned biweekly for growth measurement. In addition, thermometers were used to record temperature automatically at 1 m from the ground and at a depth of 10 cm in soil at 12 a.m. daily to evaluate the effect of temperature on fine root production. The results indicated that the combined method gave 19 and 16 % higher estimation of fine root production than decision matrix and continuous inflow method, respectively. In the combined method, using scanned images at different time intervals gave different estimates. Generally, longer time-interval images gave lower estimates: biweekly-interval images yielded an estimate that was 16 % higher than that yielded using 2-month-interval images. A 1-month time lag relationship (R 2 = 0.84, p < 0.01) between soil temperature and fine root production was found at the study site. This relationship implied that higher fine root production occurred in summer. It was concluded that the combined method gave a more accurate estimation of fine root production in forests among the three methods compared.20.
V. Brozzoli S. Crognale I. Sampedro F. Federici A. D’Annibale M. Petruccioli 《Bioresource technology》2009,100(13):3395-3402
Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l−1 NH4Cl and 3 g l−1 olive oil led to an enzyme activity of about 10 U ml−1. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml−1) when the pH was held constant at 6.5, significantly increased (18.7 U ml−1) with uncontrolled pH and was maximum (20.4 U ml−1) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml−1) and mean volumetric productivity (i.e., 123.5 U l−1 h−1). 相似文献