法夫酵母PLX-All发酵纤维素酶水解物合成虾青素*

法夫酵母（Phaffia rhodozyma)PLX-All菌株能够发酵纤维素酶水解物进行虾青素的生物合成。纤维素的酶解物主要为纤维二糖和葡萄糖,在另外添加适量其它营养物后可被法夫酵母发酵用于生长及合成虾青素。摇瓶试验结果表明,培养108h,法夫酵母的生物量可达2.3g／L,虾青素的产率达913.4g／g干细胞,虾青素体积产率为2.1mg／L。在2L罐的发酵试验中,法夫酵母的生物量可达3.23g／L（第96h）,虾青素的产率达581.4g／g干细胞,虾青素体积产率达1.88mg／L。     

法夫酵母PLX-ll发酵纤维素酶水解物合成虾青素

法夫酵母（Phaffiarhodozyma)PLX朅ll菌株能够发酵纤维素酶水解物进行虾青素的生物合成。纤维素的酶解物主要为纤维二糖和葡萄糖,在另外添加适量其它营养物后可被法夫酵母发酵用于生长及合成虾青素。摇瓶试验结果表明,培养108h,法夫酵母的生物量可达2.3g／L,虾青素的产率达913.4g／g干细胞,虾青素体积产率为2.1mg／L。在2L罐的发酵试验中,法夫酵母的生物量可达3.23g／L（第96h）,虾青素的产率达581.4g／g干细胞,虾青素体积产率达1.88mg／L。     

法夫酵母PLX—A11发酵纤维素酶水解物合成虾青素

法夫酵母（Ｐｈａｆｆｉａ ｒｈｏｄｏｚｙｍａ）ＰＬＸ－Ａ１１菌株能够发酵纤维素酶水解物进行虾青素的生物合成。纤维素的酶解物主要为纤维二糖和葡萄糖,在另外添加适量其它营养物后可被法夫酵母发酵用于生长及俣成虾青素。摇瓶试验结果表明,培养１０８ｈ,法夫酵母的生物量可达２．３ｇ／Ｌ,虾青素的产率达９１３．４μｇ／ｇ干细胞,虾青素体积产率为２．１ｍｇ／Ｌ。在２Ｌ罐的发酵试验中,法夫酵母的生物量可达３．２３ｇ     

一株法夫酵母虾青素高产菌株的生产性能

为了评价虾青素高产菌株-法夫酵母JMU-MVP14的生产性能及建立虾青素高产发酵技术,通过测定糖、生物量、虾青素产量、总类胡萝卜素产量等发酵参数,用摇瓶试验对比了法夫酵母JMU-MVP14和出发菌株的差异,用7 L罐试验对比了pH值调控方式及补料培养基成分对发酵的影响,用1 m3罐试验评估了法夫酵母JMU-MVP14高密度发酵虾青素的产量水平。摇瓶发酵结果表明,法夫酵母JMU-MVP14虾青素及总类胡萝卜素的细胞产率分别达到6.01 mg/g及10.38 mg/g;7 L罐分批发酵试验结果表明,自动流加调     

补料工艺对两株法夫酵母菌株产虾青素的影响

【目的】考察不同补料工艺对法夫酵母菌株生长和虾青素合成的影响。【方法】对法夫酵母JMU-VDL668和JMU-MVP14菌株在7 L罐中进行分批及分批补料培养; 同时, 测定发酵过程中生物量、虾青素和葡萄糖含量的变化。【结果】采用恒DO补料, 法夫酵母JMU-VDL668菌株获得的生物量最大(64.6 g/L), 是分批培养的2.2倍; 采用恒pH补料发酵, 虾青素的产量最高(20.6 mg/L), 是分批培养的1.5倍。与JMU-VDL668菌株不同, 虾青素高产菌株JMU-MVP14菌株采用恒pH补料, 获得生物量最大(48.5 g/L), 但虾青素产量大大降低(仅17.5 mg/L); 采用脉冲补料, 虾青素产量最高, 达到414.1 mg/L, 与分批发酵相比提高了200.2%; 采用恒DO补料, 生物量(38.5 g/L)和虾青素产量(403.2?mg/L)增加显著, 与分批发酵相比分别提高了133.1%和192.3%。【结论】不同补料工艺对法夫酵母菌株生产虾青素影响很大。其中, 采用恒pH补料工艺, 法夫酵母JMU-VDL668菌株可以获得最高的虾青素产量, 而采用脉冲补料工艺, 最适于法夫酵母JMU-MVP14菌株发酵生产虾青素。     

虾青素高产菌的选育

红发夫酵母（Phaffia rhodozyma)是微生物发酵法生产虾青素的优良菌株,作者采用Cs^137-γ射线重复辐照,并进行亚硝基胍（NTG）诱变处理,选育得到一株高产虾青素的红发夫酵母YB-20-28突变株,该菌株摇瓶发酵的生物量达老人家酵母YB-20-28突变株,该菌株摇瓶发酵的生物量达36.3g/L,总色素含量为1216.0μg/g,较出发菌株提高308%,虾青素产量达30.9μg/mL,是一株颇具开发潜力的虾青素高产菌株。     

利用亚硝酸钠选育法夫酵母虾青素高产菌株

以亚硝酸钠作为筛选剂选择性分离法夫酵母虾青素高产菌株。实验研究表明,在亚硝酸钠存在的情况下,法夫酵母的生长和虾青素合成量均会减少;当亚硝酸钠浓度为5000μmol/L时,法夫酵母的致死率为100%。挑取200株经过甲基磺酸乙酯（EMS）诱变后的法夫酵母,以5000μmol/L的亚硝酸钠为筛选剂摇瓶发酵后测得虾青素体积产率为正突变的菌株有87株,正突变率为43.5%。挑取其中8株进行复筛,编号为N030的菌株比出发菌株的虾青素体积产率和细胞产率分别提高了39.3%和89.3%。结果说明,亚硝酸钠可作为法夫酵母虾青素高产菌株的筛选剂,用于提高菌种的筛选效率。     

氧载体强化氧传递促进法夫酵母虾青素的合成

法夫酵母生物合成虾青素是强好氧发酵过程,溶氧水平直接影响细胞虾青素的产率。本文对虾青素的氧载体强化氧传递双液相发酵进行了研究。实验结果表明,添加豆油(作为氧载体)可提高法夫酵母发酵时的溶氧水平,促进虾青素的合成：添加豆油 0.5-5.0％ (w／v),虾青素产量随豆油添加量逐步提高,最高时达到 2.98mg/L,对照组虾青素产率为 2.50mg/L。并证明产量的提高是单位质量细胞的虾青素合成效率提高的结果。摇瓶培养时转速的高低不同,对豆油的最适添加量存在影响。较高摇瓶转速有利于豆油在培养基中分散,从而利于强化氧的传递。     

高产虾青素的红发夫酵母菌种的选育*

红发夫酵母（Phaffia　rhodozyma）是发酵法生产虾青素的优良菌株。本文采用Cs137-γ射线重复辐照,并交替进行亚硝基胍（NTG）诱变处理,选育得到一株高产虾青素的红发夫酵母YB-20-29突变株。该菌株摇瓶发酵的生物量达36.32g/L,总色素含量为1216.0μg/g,较原始菌株提高308％,虾青素产量达30.9μg/mL,是一株很有开发前景的虾青素高产菌株。     

高产虾青素的红发夫酵母菌种的选育

红发夫酵母（Phaffia rhodozyma）是发酵法生产虾青素的优良菌株。本采用Cs^137-γ射线重复辐照,并交替进行亚硝基胍（NTG）诱变处理,选育得到一株高产虾青素的红夫酵母YB-20-29突变株。该菌株摇瓶发酵的生物量达36.32g/L,总色素含量为1216.0ug/g,较原始菌株提高308%,虾青素产量达30.9ug/mL,是一株很有开发前景和虾青素高产菌株。     

Perfusion culture process plus H2O2 stimulation for efficient astaxanthin production by Xanthophyllomyces dendrorhous

A semicontinuous perfusion culture process (repeated medium renewal with cell retention) was evaluated together with batch and repeated fed-batch processes for astaxanthin production in shake-flask cultures of Xanthophyllomyces dendrorhous. The perfusion process with 25% medium renewal every 12 h for 10 days achieved a biomass density of 65.6 g/L, a volumetric astaxanthin yield of 52.5 mg/L, and an astaxanthin productivity of 4.38 mg/L-d, which were 8.4-fold, 5.6-fold, and 2.3-fold of those in the batch process, 7.8 g/L, 9.4 mg/L, and 1.88 mg/L-d, respectively. The incorporation of hydrogen peroxide (H(2)O(2)) stimulation of astaxanthin biosynthesis into the perfusion process further increased the astaxanthin yield to 58.3 mg/L and the productivity to 4.86 mg/L-d. The repeated fed-batch process with 8 g/L glucose and 4 g/L corn steep liquor fed every 12 h achieved 42.2 g/L biomass density, 36.5 mg/L astaxanthin yield, and 3.04 mg/L-d astaxanthin productivity. The lower biomass and astaxanthin productivity in the repeated fed-batch than in the perfusion process may be mostly attributed to the accumulation of inhibitory metabolites such as ethanol and acetic acid in the culture. The study shows that perfusion process plus H(2)O(2) stimulation is an effective strategy for enhanced astaxanthin production in X. dendrorhous cultures.     

法夫酵母高密度培养及虾青素的高产研究*

本文对法夫酵母Phaffia rhodozyma的不同流加培养模式进行了研究。实验结果表明,采用指数流加,虾青素产率和细胞干重具有较大值,分别达到14.52mg/l和32.56g/l;其次是恒pO2流加和恒速流加培养,虾青素产率分别达到8.89mg/l和6.70mg/l; 恒pH流加方式更有利于法夫酵母细胞的生长(14.62g/l DCW)。但是,不同流加培养模式所得的μmax和qasta具有较大的差距。恒pH、恒pO2流加培养及间歇培养有较大值,分别为0.0613 h-1、0.056 h-1、0.053 h-1;指数流加的μmax较小。间歇培养中虾青素生成比率最大,qasta=0.048×10-3h-1。     

Astaxanthin synthesis by Xanthophyllomyces dendrorhous DSM 5626 and its mutants on carrot extract medium under different illumination intensity

The effect of illumination intensity on astaxanthin synthesis by yeast Xanthophyllomyces dendrorhous DSM 5626 and its 4 mutants grown in cultures on carrot extract medium was investigated. Cell concentration, total carotenoid and astaxanthin yields were assessed in obtained cultures. Collected data were used to construct regression models describing the effect of illumination intensity on controlled parameters. Maximum cellular (0.44–0.46 g/kg of dry cell weight) and volumetric yields (2.3–2.4 mg/L) of the pigment were observed for mutants 10BE and 34B at 600 lx, as well as for mutant 26UV at 1000 lx. The highest yield of astaxanthin for the parental strain was obtained in culture at illumination of 1000 lx (0.29 g/kg of dry cell weight and 1.51 mg/L). The values of illumination determined on the basis of constructed regression models for individual yeast strains, at which astaxanthin synthesis should be the most efficient, remained within the range of 660–1000 lx.     

Optimization of Phaffia rhodozyma continuous culture through response surface methodology

Response surface methodology was applied to optimize the growth of the yeast Phaffia rhodozyma in continuous fermentation using peat hydrolysates as substrate. A second-order, complete, factorial design of the experiments was used to develop empirical models providing a quantitative interpretation of the relationships between the two variables studied, dilution rate and pH. Maximum biomass concentration in the fermentor was obtained by employing the following predicted optimum fermentation conditions: a dilution rate of 0.017/h and a pH level of 7.19. A verification experiment, conducted at previously optimized conditions for maximum biomass volumetric productivity (a dilution rate of 0.022/h, and a pH level of 6.90), produced values for biomass concentration, residual substrate concentration, biomass yield, and biomass volumetric productivity that were very close to the predicted values, indicating the reliability of the empirical model. The concentration of the pigment astaxanthin produced by the yeast under the optimized growth conditions was found to be 544 mg astaxanthin/kg dry cell biomass.     

Modeling of Xanthophyllomyces dendrorhous growth on glucose and overflow metabolism in batch and fed-batch cultures for astaxanthin production

An astaxanthin-producing yeast Xanthophyllomyces dendrorhous ENM5 was cultivated in a liquid medium containing 50 g/L glucose as the major carbon source in stirred fermentors (1.5-L working volume) in fully aerobic conditions. Ethanol was produced during the exponential growth phase as a result of overflow metabolism or fermentative catabolism of glucose by yeast cells. After accumulating to a peak of 3.5 g/L, the ethanol was consumed by yeast cells as a carbon source when glucose in the culture was nearly exhausted. High initial glucose concentrations and ethanol accumulation in the culture had inhibitory effects on cell growth. Astaxanthin production was partially associated with cell growth. Based on these culture characteristics, we constructed a modified Monod kinetic model incorporating substrate (glucose) and product (ethanol) inhibition to describe the relationship of cell growth rate with glucose and ethanol concentrations. This kinetic model, coupled with the Luedeking-Piret equation for the astaxanthin production, gave satisfactory prediction of the biomass production, glucose consumption, ethanol formation and consumption, and astaxanthin production in batch cultures over 25-75 g/L glucose concentration ranges. The model was also applied to fed-batch cultures to predict the optimum feeding scheme (feeding glucose and corn steep liquor) for astaxanthin production, leading to a high volumetric yield (28.6 mg/L) and a high productivity (5.36 mg/L/day).     

一株产虾青素的黄杆菌CF—60的研究

从土壤中分离到一株黄杆菌（Ｆｌａｖｏｂａｃｔｅｒｉｕｍｓｐｐ）Ｃ(Ｆ－６０),该菌的生长需Ｍｇ２＋存在,ＭｇＳＯ４·７Ｈ２Ｏ的最适浓度为０．２％;蛋白胨是该菌株生长的最好氮源,它不能利用无机氮。种龄超过９６ｈ的菌体不能在新鲜培养基中生长。经５４ｈ的２Ｌ恒化器发酵,生物量达６．８ｇ／Ｌ,色素产量为１０．６ｍｇ／Ｌ。该菌产生的类胡萝卜素成分简单,主要成分的含量为９０．３％,该成分经初步鉴定是分子结构中含有羰基和羟基的虾青素。     

不同碳源对小球藻Chlorella zofingiensis异养产虾青素的影响

研究了葡萄糖、蔗糖和果糖对小球藻(Chlorella zofingiensis)异养生长及产虾青素的影响,结果表明,在糖浓度为20g/L时,细胞生长较快,但干重较小,虾青素含量较低;在糖浓度为50g/L时,细胞生长较慢,但干重较大,虾青素含量较高。3种碳源中蔗糖和葡萄糖效果较好,在蔗糖浓度为50g/L时,虾青素含量和产量分别达到0.94mg/g和9.61mg/L。     

Growth kinetics and astaxanthin production of Phaffia rhodozyma on glycerol as a carbon source during batch fermentation

Glycerol was studied as a substrate for astaxanthin by Phaffia rhodozyma PR 190. With co-utilisation of yeast extract and peptone, the maximum specific growth rate was 0.24 ± 0.02 h–1. Astaxanthin percentage in total pigment is constant (0.78 mg/g) and its yield from glycerol is always 0.97 mg/g. The yield of biomass from glycerol alone is 0.50 ± 0.02 g/g. The specific rate of astaxanthin production versus the cell growth rate reached a maximum for an optimal specific growth rate of 0.075 h–1. For this optimal value, the maximum specific astaxanthin production rate is 0.09 ± 0.01 mg/g.h. The best astaxanthin results were : 33.7 mg/l, 0.2 mg/l.h and 1.8 mg/g yeast after a fermentation term of 168 hours. Our results suggest a strategy of astaxanthin production in fed batch culture or chemostat at a growth rate of 0.075 h–1. © Rapid Science Ltd. 1998