Kojic acid production in an airlift bioreactor using partially hydrolyzed raw corn starch

In a culture of Aspergillus oryzae MK-107-39 in a 3-l airlift bioreactor, kojic acid was not produced when glucose/wheat germ medium (GM1) was used. However, when a jar fermentor was used, the kojic acid yield was high. A suitable medium for culture in an airlift bioreactor consisting of partially hydrolyzed corn starch and a small amount of corn steep liquor (CSL) (SM1) was selected. In the cultivation in the airlift bioreactor using SM1, nearly 40 g/l of kojic acid was produced, which was the same as the amount produced in the jar fermentor containing GM1. The optimum aeration rate for the airlift bioreactor was 2.0 vvm (0.66 cm/s of superficial linear velocity (Vs)). The cost of SM1 using the airlift bioreactor was reduced to 40% that of GM1 using the jar fermentor. Furthermore, the energy cost of kojic acid production using SM1 in the airlift bioreactor was less than one-fourth of that for the jar fermentor using GM1.     

Continuous production of wine vinegar in bubble column reactors of up to 60-litre capacity

This paper describes the development of a continuous acetic acid fermentation process for the production of wine vinegar in bubble column reactors of up to 60 l capacity. To determine appropriate fermentation conditions a study of the influence of residual ethanol concentration, inlet flow rate and aeration was carried out using a 6-l laboratory reactor, white table wine as fermentation medium, a temperature of 30 °C and an air flow rate of 0.125 min^-1 (vvm). The concentration of acetic acid obtained in the continuous wine vinegar production ranged from 91 g/l at 28.6 ml/h to 28 g/l at 154.1 ml/h by increasing the inlet flow rate. As expected, the biomass decreased as well, from 208 mg/l to 106 mg/l. The maximum acetification rate was observed in the range 85-110 ml/h, corresponding to a value of about 1.1 g/l/h. A further increase in the flow rate produced a slight decrease in the acetification rate. Best yields, between 94.5 and 94.7%, were obtained in the flow rate range of 60-75 ml/h. The acetification rate was improved only by about 10% by increasing the aeration from 0.125 to 0.250 min^-1. The continuous wine vinegar production was scaled up from the laboratory fermentor to a 60-l pilot acetator. During the steady state (residential time >6), with an inlet flow rate of 950 ml/h, temperature of 30 °C and aeration of 0.250 min^-1, the following parameters were obtained: acetic acid concentration 72 g/l, overall productivity 1.41 g/l/h and yield 94.2%.     

微生物转化香草酸生产香草醛

筛选到 1株可以转化香草酸生成香草醛的朱红密孔菌 (Pycnoporuscinnabarinus)SW 0 2 0 3 ,通过培养基和转化条件的优化后 ,可以将 1 682g/L的香草酸转化生成 0 875 g/L香草醛 ,摩尔转化率为 5 7 5 %。在 2 5L罐上进行放大试验 ,香草醛产量为 0 81 8g/L ,摩尔转化率为 5 3 7%。对转化液中的产物进行提取 ,得到纯度为 95 %的香草醛结晶 ,提取收率为63 9%。     

Effect of aeration and agitation on growth rate of Thermus thermophilus in batch mode

The growth kinetics of Thermus thermophilus HB27 was investigated in rich medium (Thermus medium) under batch cultivation at 65 degrees C in 3-l fermentors. The growth and oxygen consumption rates were highly dependent on the aeration and agitation rates. Volumetric mass transfer coefficient (K(L)a, h(-1)) and hence oxygen transfer rate (OTR, mol m (-3) h(-1)) into the fermentation broth increased with increased aeration and/or agitation rates. A K(L)a and OTR of 175.4 h(-1) and 31.7 mol m(-3)h(-1) respectively, corresponding to 500 rpm agitation and 2 vvm aeration with a mixture of air and oxygen, were required to avoid oxygen limitation. The maximum growth rate (mu(max), h(-1)), doubling time (t(D), h), and dry cell weight determined for T. thermophilus HB27 growing under these conditions were 0.27 h(-1), 2.67 h, and 3g/l respectively. This cell yield is higher than any previously published reports for growth of Thermus cultures, including studies that employed pressurized bioreactors.     

Wastewater treatment with bacteria immobilized onto a ceramic carrier in an aerated system

Biological treatment of the wastewater discharged from a food processing factory was continuously carried out in a packed bed bioreactor under aerobic conditions. The bacterium isolated from the wastewater was immobilized onto a new type of ceramic carrier by a vacuum method and high numbers of bacteria were colonized onto the carrier (2.9 x 10(9) cfu/g of dry ceramic carrier). The effect of the hydraulic retention time (HRT) and aeration rate on the removal of the chemical oxygen demand (COD) was investigated. The system was able on average to remove more than 82% of the influent COD during 160 d of operation and more than 87% of the influent COD on average was removed when the HRT was 30.17 h and the aeration rate was 2.0 vvm. Aeration rates in the range of 0.4 to 2.0 vvm do not affect the COD removal efficiency.     

搅拌转速和通气量对灰树花深层发酵培养的影响

采用15L搅拌式发酵罐研究灰树花的扩大培养条件，主要考察了搅拌转速和通气量对灰树花菌丝量、胞外多糖产率、溶氧、菌丝形态以及发酵液粘度的影响．研究结果表明搅拌转速和通气量能显著影响灰树花的生长，并且通过对灰树花菌丝球的大小、形态以及胞外多糖的分泌影响发酵液的粘度．并得出较为优化的培养条件为：温度25℃，通气量0．75vvm，搅拌转速80r／min，装液量60％；在此条件下，发酵10d后，灰树花菌丝体最大得率为22．95g／L，胞外多糖的得率为1．521g／L。     

Optimization of β-carotene production by a mutant of the lactose-positive yeast Rhodotorula acheniorum from whey ultrafiltrate

The present work investigated on carotenogenesis with high β-carotene content by a new isolated high-activity strain-producer Rhodotorula acheniorum mutant MRN in cheese whey ultrafiltrate. After a serial of UV, ethymethanesurfonate (EMS), and nitrosoguanidine (NTG) mutagenesis, a mutant named MRN of the red lactose-positive yeast strain R. acheniorum was obtained. Then, the effects of different growth medium factors on carotenoid production by this mutant at batch-scale level were identified and optimized by means of response surface methodology (RSM) in order to achieve high-level production of β-carotene. The optimum conditions required to achieve the highest level of β-carotene (262.12±1.01 mg/L) were determined as follows: whey ultrafiltrate (WU) lactose concentration 55 g/L, pH 5.85, ammonium sulfate concentration 3.5 g/L, temperature 23°C, and aeration rate 1.56 vvm. The medium optimization resulted in a 6.45-fold increase in volumetric production (262.12±1.01 mg/L) and a 4.62-fold increase in the cellular accumulation (10.69±0.19 mg/g) of β-carotene.     

Process development for mycelial growth and polysaccharide production in Tricholoma matsutake liquid culture

In this study, the effects of agitation and aeration on mycelial growth and exo-polysaccharide production were examined in batch cultures of Tricholoma matsutake. Agitation was varied from 100 to 300 rpm and aeration was varied from 0.5 to 1.5 vvm. Mycelial growth was 21.87 g/l at 150 rpm, and exo-polysaccharide production was 8.79 g/l at 1.5 vvm. When we analyzed the polysaccharide extractions from the cultured mycelium and the culture broth of T. matsutake, 1.4 g of crude polysaccharide was found per 100 g of dried weight in the cultured mycelium, and 1.47 g/l of polysaccharides was found in the culture broth. In addition, the amounts of β-Glucan in the soluble polysaccharide fractions of the cultured mycelium and culture broth were 75.4% and 83.6%, respectively. The cultured mycelium and the culture broth contained a higher amount of β-Glucan than that of the fruiting body.     

Medium optimization using mathematical statistics for production of β-Carotene by Blakeslea trispora and fermenting process regulation

Optimization of medium components for enhancement of β-carotene production by Blakeslea trispora was achieved using mathematical statistics. Optimum concentrations of components using Plackett-Burman design and response surface methodology (RSM) were d-glucose 7.16%, wheat bran extract 4.08%, MgSO₄ 0.04%, soybean oil 3%, thiamine 0.01%, soybean meal 1%, and KH₂PO₄ 0.2%. Maximum production of β-carotene using optimized medium was 643 mg/L in a shake flask. A predicted value of 669 mg/L was based on results of an RSM regression. The optimum aeration rate of 1.5 vvm produced 866 mg/L and the optimum agitation speed of 100 rpm produced 975 mg/L of β-carotene. The t of a quadratic model using regression derived coefficients was significant. Maximum production of β-carotene using the optimized medium in a stirred tank reactor (10 L) at an optimal aeration rate and an optimum agitation speed with addition of 0.1%(v/v) β-ionone was 1,357 mg/L.     

Production of extracellular bifidogenic growth stimulator (BGS) from Propionibacterium shermanii using a bioreactor system with a microfiltration module and an on-line controller for lactic acid concentration

Production of a bifidogenic growth stimulator (BGS) by Propionibacterium freudenreichii subsp. shermanii (Propionibacterium shermanii) using lactic acid as a carbon source was investigated using different cultivation methods. When a continuous bioreactor system with a filtration device was used at a dilution rate of 0.075 h(-1), the average BGS concentration was 2.4 mg/l, which corresponds to a BGS productivity per cultivation time of 1.8 x 10(-1) mg x l(-1) x h(-1). The BGS productivity per cultivation time in continuous cultivation with filtration was 1.9-fold that (9.4 x 10(-2) mg x l(-1).h(-1)) in a conventional batch cultivation. In fed-batch cultivation with feed-back control using an on-line lactic acid controller with a lactic acid biosensor, it was possible to prevent substrate inhibition by maintaining the lactic acid concentration in culture broth low at 3.3 g/l, and an enhanced BGS production (31 mg/l) was successfully attained. The BGS productivity per cultivation time (2.1x10(-1) mg x l(-1) x h(-1)) in the fed-batch cultivation with feed-back control was 2.2-fold that in the conventional batch cultivation. A new bioreactor system was developed by coupling a continuous bioreactor system with a filtration device to an on-line lactic acid controller. Using the new bioreactor system, we produced BGS continuously at a high level of 47 mg/l. The BGS productivities per cultivation time (3.5 mg.l(-1) x h(-1)) and the total volume of medium used (1.7 x 10(-1) mg x l(-1) x h(-1)) obtained in the new bioreactor system were 37-fold and 2.1-fold those in the conventional batch cultivation, respectively. These results described above clearly demonstrate the positive effects of both the continuous filtration for removal of metabolites (propionic and acetic acids) inhibitory to cell growth and feed-back control of lactic acid concentration in the culture broth on BGS production by P. shermanii. This paper is the first report on BGS production by the propionic acid bacterium using lactic acid as a carbon source.     

Optimization and scale-up of L-lactic acid fermentation by mutant strain Rhizopus sp. MK-96-1196 in airlift bioreactors

We determined the optimum culture conditions such as inoculum size, initial starch concentration, pH during the fermentation and aeration rate for L-lactic acid production by Rhizopus sp. MK-96-1196 in a 3-l airlift bioreactor. More than 90 g/l of L-lactic acid was produced from only partially enzymatically hydrolyzed corn starch with a production rate of 2.6 g/l/h and a product yield of 87% based on the starch consumed under the optimum conditions in the 3-l airlift bioreactor. Scale-up from the 3-l to a 100-l airlift bioreactor for L-lactic acid fermentation was carried out using V(s)(cm/s) as a scale-up criterion. The production rates and yields of L-lactic acid in both bioreactors appeared to be fairly well correlated with k(L)a (1/h).     

Evaluation of the conditions of carotenoids production in a synthetic medium by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor

This work studied the cultivation conditions for the production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor. A Plackett–Burman design was used for the screening of the most important factors, followed by a complete second order design, to maximise the concentration of total carotenoids. The maximum concentration of 3425.9 μg L^?1 of total carotenoids was obtained in a medium containing 80 g L^?1 glucose, 15 g L^?1 peptone and 5 g L^?1 malt extract, with an aeration rate 1.5 vvm, 180 r.p.m., 25 °C and an initial pH of 4.0. Fermentation kinetics showed that the maximum concentration of total carotenoids was reached after 90 h of fermentation. Carotenoid bio‐production was partially associated with cell growth. The specific carotenoid production (Y_P/X) was 238 μg carotenoids/g cells, whereas Y_P/S (substrate to product yield) was 41.3 μg g^?1. The specific growth rate (μ_x) was 0.045 h^?1. The highest cell and total carotenoid productivity were 0.19 g L^?1 h^?1 and 56.9 μg L^?1 h^?1, respectively.     

Continuous plant cell perfusion culture: bioreactor characterization and secreted enzyme production

Culture perfusion is widely practiced in mammalian cell processes to enhance secreted antibody production. Here, we report the development of an efficient continuous perfusion process for the cultivation of plant cell suspensions. The key to this process is a perfusion bioreactor that incorporates an annular settling zone into a stirred-tank bioreactor to achieve continuous cell/medium separation via gravitational sedimentation. From washout experiments, we found that under typical operating conditions (e.g., 200 rpm and 0.3 vvm) the liquid phase in the entire perfusion bioreactor was homogeneous despite the presence of the cylindrical baffle. Using secreted acid phosphatase (APase) produced in Anchusa officinalis cell culture as a model we have studied the perfusion cultures under complete or partial cell retention. The perfusion culture was operated under phosphate limitation to stimulate APase production. Successful operation of the perfusion process over four weeks has been achieved in this work. When A. officinalis cells were grown in the perfusion reactor and perfused at up to 0.4 vvd with complete cell retention, a cell dry weight exceeding 20 g/l could be achieved while secreted APase productivity leveled off at approximately 300 units/l/d. The culture became extremely dense with the maximum packed cell volume (PCV) surpassing 70%. In comparison, the maximum cell dry weight and overall secreted APase productivity in a typical batch culture were 10-12 g/l and 100-150 units/l/d, respectively. Operation of the perfusion culture under extremely high PCV for a prolonged period, however, led to declined oxygen uptake and reduced viability. Subsequently, cell removal via a bleed stream at up to 0.11 vvd was tested and shown to stabilize the culture at a PCV below 60%. With culture bleeding, both specific oxygen uptake rate and viability were shown to increase. This also led to a higher cell dry weight exceeding 25 g/l, and further improvement of secreted APase productivity that reached a plateau fluctuating around 490 units/l/d.     

真菌固定床反应器发酵L(+)-乳酸的初步研究

研究了在5L发酵罐中采用纤维床固定化技术发酵生产乳酸过程不同溶氧量和固定化面积对乳酸发酵过程的影响,测定了不同固定化面积发酵过程中的氧传质系数。在通气率为1.0vvm和固定化面积为400cm2条件下,产酸速率为2.13g/(L·h),发酵液中乳酸最终浓度为73.1g/L,L(+)-乳酸光学纯度为98.9%,证明提高发酵过程中菌体层内部溶氧传质系数有助于增加乳酸产率。乳酸发酵液浊度0.49NTU,显著改善发酵液的流体力学条件,为应用膜生物反应器技术连续发酵生产乳酸奠定了基础。     

Application of a complete factorial design for the production of zeaxanthin by Flavobacterium sp

Utilizing a four-liter fermentor and applying a complete factorial design 2(3), the combined effects of agitation speed, aeration rate, and corn steep liquor concentration on zeaxanthin production by Flavobacterium sp. were studied. Maximum growth and production of total carotenoids and zeaxanthin were obtained at 600 rpm, 2 vvm and 4.6% corn steep liquor. Under these conditions, zeaxanthin represented 86% of the total carotenoids produced. Lower values of the variables studied resulted in lower growth, volumetric production of zeaxanthin and total carotenoids, and favored the formation of other carotenoids such as beta-carotene and canthaxanthin. The positive effects on growth and total carotenoids and zeaxanthin formation were in a large extent due to the interaction of agitation/corn steep liquor. However, aeration also had a positive effect on growth.     

两株丝状真菌联合转化生产香草醛的研究

本文对黑曲霉(Aspergillus niger)CGMCC0774和朱红密孔菌(Pycnoporus cinnabarinus)CGMCC1115联合转化来源于米糠油脚的阿魏酸生产香草醛进行研究。通过60Co诱变黑曲霉菌株,得到耐较高底物浓度且不降解产物的突变株产香草酸2.48g/L;其转化液经过滤,调pH至5.0,补加5g/L葡萄糖,灭菌处理后,作为朱红密孔菌离菌体转化的底物溶液进行转化,得到1.07g/L香草醛。     

Production of podophyllotoxin by plant cell cultures of Podophyllum hexandrum in bioreactor

Submerged cultivation of Podophyllum hexandrum for the production of podophyllotoxin was carried out in a 3l stirred tank bioreactor fitted with a low-shear Setric impeller. The specific requirements of the medium, such as carbon source (sugar) and light, were established for the growth of and podophyllotoxin production by P. hexandrum in suspension cultures. Substitution of sucrose by glucose resulted in higher growth and podophyllotoxin production. The biosynthesis of podophyllotoxin was favored when plant cells were cultivated in the dark. An agitation speed of 100 rpm was sufficient to mix the culture broth in the bioreactor without causing any significant cell damage. Biomass and podophyllotoxin accumulation in 3 l bioreactor under batch growth conditions were 6.5 g/l and 4.26 mg/l, respectively, in 22 d. This resulted in an overall podophyllotoxin productivity of 0.19 mg/(l.d), which represented an increase of 27% in comparison to its productivity in a shake flask. Podophyllotoxin production was found to be a combined growth-associated and non-growth associated process.     

微生物转化阿魏酸生产香兰素的研究

对朱红秘孔菌的生长和香兰素的生成两个阶段分别采用高密度培养基和普通培养基进行培养发酵,并通过对香兰素生成阶段的发酵条件的优化后,可生成0．403g／L的香兰素,相应的摩尔转化率为28．8％。     

Effects of pH and dissolved oxygen on cellulose production by Acetobacter xylinum BRC5 in agitated culture

Acetobacter xylinum BRC5 was cultivated in a jar fermentor using glucose as the sole carbon source. Strain BRC5 oxidized almost all of the glucose to gluconic acid; thereafter, it biosynthesized cellulose by utilizing gluconic acid accumulated in the broth. The optimal pH for metabolizing glucose to gluconic acid was 4.0, while a pH of 5.5 was preferred for cell growth and cellulose production from the accumulated gluconic acid in the medium. Shifting the pH from 4.0 to 5.5 during the cellulose production phase in batch cultures improved cellulose production and reduced the total fermentation time, compared to batch cultures at constant pH. In constant fed-batch culture, 10 g/l of cellulose was obtained from 40 g/l of glucose, a yield which was approximately 2-fold higher than in batch culture with the same initial glucose concentration, even without control of the level of dissolved oxygen. The highest cellulose yield was obtained in fed-batch cultures in which the dissolved oxygen concentration was controlled at 10% saturation. Control of pH and dissolved oxygen to optimal levels was effective for improving the production rate and yield of cellulose, to achieve a high cellulose productivity of 0.3 g cellulose/l x h. Approximately 15 g/l of cellulose was considered to be the highest yield obtainable using conventional fermentors because the culture broth then became too viscous to allow satisfactory aeration.     

Performance of a partially packed charcoal pellet bioreactor for acetic acid fermentation

The performance of a partially packed charcoal pellet bioreactor was compared to that of a fully packed bioreactor for aerobic acetic acid production. In the fully packed charcoal pellet bioreactor, it was considered that the shortening of an actual retention time of the culture broth limited the bioreactor performance under high dilution rate and high aeration conditions. By reducing the filling ratio of charcoal pellets to 44%, which increased the actual retention time of the culture broth, the maximum productivity increased from 3.9 g/l/h in the fully packed bed bioreactor to 5.7 g/l/h in the partially packed bioreactor without affecting the operational stability.