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。     

温度对雨生红球藻(Haematococcus pluvialis)生物量和虾青素产量的影响

在用环形培养池模拟系统培养雨生红球藻的过程中,研究了温度对雨生红球藻生物量及虾青素产量的影响。结果表明,在15～25℃的范围内,不同温度下雨生红球藻生物量和虾青素含量及产量都经历了一个上升—最高—下降的过程。25℃与22℃时红球藻的虾青素产量、虾青素含量(干重)均显著高于其他温度的(P<0.01),但两者间差异不显著(P>0.05)。15℃时,红球藻生物量、虾青素含量和虾青素产量均最低,分别为1.4g、0.54%和2.49mg/L;25℃时,红球藻生物量和虾青素产量最高,分别为2.68g和13.53mg/L;22℃时,虾青素含量最高,为1.52%。     

基于动力学模型的法夫酵母发酵生产虾青素的补料策略优化

对法夫酵母的不同补料发酵方式进行了研究.基于底物抑制模型,提出了一种优化的两阶段补料策略,用于法夫酵母产虾青素的高密度发酵.在发酵的延迟期和对数生长期早期,糖浓度控制在25 g/L左右,在此条件下,生物量可以达到最大,且时间缩短.在对数生长期后期及稳定期,糖浓度控制在5 g/L,虾青素的合成时间可以有效延长.与传统的补料方式相比,采用此补料策略取得了较好的发酵效果.发酵终点细胞干重达到23.8g/L,虾青素产量达到29.05 mg/L,分别比分批发酵提高了52.8%和109%.     

黄腐酸对雨生红球藻虾青素的积累和CHY基因表达量影响

实验以雨生红球藻Haematococcus pluvialis LUGU为对象,研究了不同浓度的黄腐酸对微藻细胞生长、虾青素积累以及β-胡萝卜素羟化酶(CHY)基因表达量的影响。结果表明,FA浓度为5 mg/L,藻细胞生物量产率达到了79.39 mg/(L·d),虾青素产量达到了20.82 mg/L,分别比对照组提高了4.25%和86.89%;FA浓度为10 mg/L,藻细胞的生物量产率和虾青素产量分别比对照组提高了5.44%和9.78%。RT-PCR分析显示,虾青素合成的关键基因CHY的表达受FA的诱导,当添加5和10 mg/L的黄腐酸时,CHY基因最大的表达量分别为对照的18.1倍和7.3倍,当添加20 mg/L的黄腐酸时CHY基因的最大的表达量仅为对照的3.2倍,FA诱导下的雨生红球藻虾青素的积累含量和CHY基因表达量呈正相关。实验表明,适当浓度的黄腐酸不仅能够显著提高虾青素合成关键酶基因CHY的表达水平,并且明显促进了藻细胞内虾青素的积累,因此黄腐酸可作为虾青素生产的一种有效诱导子。     

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-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-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。     

碳氮比和光强对小球藻合成虾青素的影响

本文研究了碳氮比和光强对小球藻Chlorella zofingiensis合成虾青素的影响。结果表明, 随着碳氮比的增加, 虾青素含量提高, 但是过高的碳氮比限制藻细胞的生长。当碳氮比为 133 时, 虾青素的产量最大, 达到 9.19 mg/L。高光诱导在高碳氮比基础上进一步提高虾青素的含量及产量。200 mmol/m2﹒s 光强在没有严重影响小球藻生长的同时, 明显提高虾青素含量, 最大虾青素产量达到 12.52 mg/L。文章对不同诱导条件下虾青素的合成机理以及利用异养自养相结合的方法提高虾青素产量的可能性进行了讨论。     

几种营养物质对烟草细胞生长和CoQ_(10)含量的影响

研究了蔗糖、KH2PO4、NH 4/NO3比对烟草细胞生长和CoQ10含量的影响,结果表明,当蔗糖浓度为30g/L时,CoQ10总量最高,此时细胞产量、CoQ10含量和总量分别为198g/L、4147μg/g(dwt)、8212μg/L。在上述蔗糖浓度下,当KH2PO4起始浓度为340mg/L、NH 4/NO-3为12时,细胞产量、CoQ10含量和总量分别最高,高于此比例时有利于CoQ10形成,但不利于细胞生长。     

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。     

Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column

A gradient reversed-phase high-performance liquid chromatography (HPLC) method using a C30 column was developed for the simultaneous determination of astaxanthin, astaxanthin monoesters and astaxanthin diesters in the green algae Chlorococcum sp., Chlorella zofingiensis, Haematococcus pluvialis and the mutant E1, which was obtained from the mutagenesis of H. pluvialis by exposure to UV-irradiation and ethyl methanesulphonate (EMS) with subsequent screening using nicotine. The results showed that the contents of total astaxanthins including free astaxanthin and astaxanthin esters ranged from 1.4 to 30.9 mg/g dry biomass in these green algae. The lower total astaxanthin levels (< 2 mg/g dry biomass) were detected in the green algae Chlorococcum sp. and C. zofingiensis. The higher total astaxanthin levels (>16 mg/g dry biomass) were found in the green alga H. pluvialis and its mutant E1. It is notable that the mutant E1 is found to have considerably higher amounts of total astaxanthin (30.9 mg/g) as compared to the wild strain of H. pluvialis (16.1 mg/g). This indicates that UV-irradiation and EMS compound mutagenesis with subsequent screening using nicotine is an effective method for breeding of a high-producing astaxanthin strain of H. pluvialis. In addition, the green alga C. zofingiensis had a remarkably higher percentage of astaxanthin diesters (76.3% of total astaxanthins) and a remarkably lower percentage of astaxanthin monoesters (18.0% of total astaxanthins) in comparison with H. pluvialis (35.5% for diesters and 60.9% for monoesters), the mutant E1 (49.1% and 48.1%) and Chlorococcum sp. (18.0% and 58.6%). Supported by the Frontier Research Grant of the SCSIO, the Hundred Talents program of Chinese Academy of Sciences, and National Natural Sciences of China projects (Grant No. 40776087)     

Growth-associated biosynthesis of astaxanthin in heterotrophic Chlorella zofingiensis (Chlorophyta)

The green microalga Chlorella zofingiensis can produce the ketocarotenoid astaxanthin under heterotrophic culture conditions. Here we report the growth-associated biosynthesis of astaxanthin in this biotechnologically important alga. With glucose as sole carbon and energy source, C. zofinginesis grew fast in the dark with rapid exhaustion of nitrogen and carbon sources from media, leading to a high specific growth rate (0.034 h⁻¹). Cultures started at a cell concentration of about 3.4 × 10⁹ cells l⁻¹ reached, after 6 days, standing biomass values of 1.6 × 10¹¹ cells or 8.5 g dry weight l⁻¹. Surprisingly, the biosynthesis of astaxanthin was found to start at early exponential phase, independent of cessation of cell division. A general trend was observed that the culture conditions benefiting cell growth also benefited astaxanthin accumulation, indicating that astaxanthin was a growth-associated product in this alga. The maximum cell dry biomass and astaxanthin yield were 11.75 g l⁻¹ and 11.14 mg l⁻¹ (about 1 mg g⁻¹), simultaneously obtained in the fed-batch culture with a combined glucose–nitrate mixture addition, which were the highest ever reported in dark-heterotrophic algal cultures. The possible reasons why dark-heterotrophic C. zofingiensis could produce astaxanthin during the course of cell growth were discussed.     

Comparison of the accumulation of astaxanthin in Haematococcus pluvialis and other green microalgae under N-starvation and high light conditions

Haematococcus pluvialis gave the highest astaxanthin accumulation rate (2.7 mg l^–1 day^–1) and total astaxanthin content ( 22.7 mg g^–1 biomass). Astaxanthin accumulation in Neochloris wimmeri, Protosiphon botryoides, Scotiellopsis oocystiformis, Chorella zofingiensis and Scenedesmus vacuolatus was, respectively, 19.2, 14.3, 10.9, 6.8 and 2.7 mg astaxanthin g^–1 biomass, respectively.     

Utilization of cane molasses towards cost-saving astaxanthin production by a Chlorella zofingiensis mutant

The aim of the present study was to survey the growth and astaxanthin production of E17, an astaxanthin-rich mutant of Chlorella zofingiensis, through feeding the low-cost carbon source cane molasses. In heterotrophic batch cultivation, E17 fed with pretreated molasses achieved biomass (1.79 g L^?1 day^?1) and astaxanthin (1.99 mg L^?1 day^?1) productivities comparable to those with glucose, which were about 2- and 2.8-fold of those fed with untreated molasses, respectively. Molasses-induced astaxanthin accumulation may be attributed to the elicited expression of carotenogenic genes, in particular the genes specifically responsible for the ketolation and hydroxylation of β-carotene to form astaxanthin. A two-stage fed-batch strategy was employed to grow E17 and induce astaxathin accumulation, resulting in 45.6 g L^?1 biomass and 56.1 mg L^?1 astaxanthin, the highest volumetric astaxanthin yield ever reported for this alga. In addition, the astaxanthin production by E17 was tested with a semi-continuous culture method, where the directly diluted raw molasses (giving 5 g L^?1 sugar) was used as the carbon source. Little growth inhibition of E17 was observed in the semi-continuous culture with a biomass productivity of 1.33 g L^?1 day^?1 and an astaxanthin productivity of 0.83 mg L^?1 day^?1. The mixotrophic semi-continuous cultures enhanced the biomass and astaxanthin productivities by 29.3 % and 42.2 %, respectively. This study highlights the potential of using the industrially cheap cane molasses towards large-scale cost-saving production of the high-value ketocarotenoid astaxanthin.     

The Effect of Salt Stress on the Production of Canthaxanthin and Astaxanthin by Chlorella zofingiensis Grown Under Limited Light Intensity

The fresh water green microalga Chlorella zofingiensisis known to accumulate ketocarotenoids – primarily astaxanthin but also canthaxanthin – when grown under stress conditions of high light irradiance and low nitrogen. We found that salt stress can replace light stress with respect to inducing carotenoid production: cells of C. zofingiensis grown under low light irradiance and subjected to salt and low nitrogen stress accumulated higher amounts of total secondary carotenoids than those growing under high light and low nitrogen stress. Furthermore, C. zofingiensis growing under conditions of salt stress and low light accumulated higher amounts of canthaxanthin than astaxanthin. It is suggested that for canthaxanthin accumulation under salt stress, light is not a limiting factor, but for astaxanthin accumulation high light irradiance is mandatory. These results may be applied in the future for the commercial production of canthaxanthin by C. zofingiensis in systems in which light availability is poor.     

Cellulose production from<Emphasis Type="Italic">Gluconobacter oxydans</Emphasis> TQ-B2

Gluconobacter oxydans that produces the cellulose was isolated. In order to confirm the chemical features of cellulose, various spectrophtometeric analysis were carried out using electron microscopy, X-ray diffractogram, and CP/MAS¹³C NMR. The purified cellulose was found to be identical to that ofAcetobacter xylinum. For effective production of cellulose, the various carbon and nitrogen sources, mixture of calcium and magnesium ions, and biotin concentration were investigated in flask cultures. Among the various carbon sources, glucose and sucrose were found to be best for the production of cellulose, with maximum concentration of 2.41 g/L obtained when a mixture of 10 g/L of each glucose and sucrose were used. With regard to the nitrogen sources, when 20 g/L of yeast extract was used, the maximum concentration of bacterial cellulose was reached. The concentration of cellulose was increased with mixture of 2 mM of each Ca²⁺ and Mg²⁺. The optimum biotin concentration for the production of cellulose was in the range of 15 to 20 mg/L. At higher biotin concentration (25–35 mg/L), the bacterial cellulose production was lower.     

Effect of light supply and carbon source on cell growth and cellular composition of a newly isolated microalga Chlorella vulgaris ESP‐31

Light supply is one of the most important factors affecting autotrophic growth of microalgae. This study investigated the effect of the type and light intensity of artificial light sources on the cell growth of an indigenous microalga Chlorella vulgaris ESP‐31 obtained from southern Taiwan. In addition, a dissolved inorganic carbon source (i.e. sodium bicarbonate) was used to improve the biomass production of strain ESP‐31. The results show that a new fluorescent light source (TL5) was effective in indoor cultivation of microalgae. Better overall productivity of 0.029 g dry cell weight/L‐d was obtained when using TL5 lamps as the light source with a light intensity of 9 W/m². A carbon source (sodium bicarbonate) concentration of 1000 mg/L was found to be optimal for the growth of strain ESP‐31 in terms of both biomass production and carbon source utilization. Under the optimal growth conditions, the resulting microalgal biomass consisted of 25–30% protein, 6–10% carbohydrate, and 30–40% lipid.     

盐单胞菌利用短链脂肪酸合成聚羟基脂肪酸酯

盐单胞菌(Halomonas)能够利用多种底物为碳源生长,由于其能在高盐条件下进行不灭菌的开放发酵,已被开发用作下一代生物技术的底盘细胞。包括乙酸、丙酸和丁酸在内的短链挥发性脂肪酸能够以生物质为原料制备,有望成为用于微生物发酵的新型碳源。利用10-50 g/L浓度的丁酸为碳源对Halomonassp.TD01和TD08进行摇瓶培养,结果发现Halomonassp.TD01和TD08能够有效利用丁酸合成聚-3-羟基丁酸酯,20 g/L丁酸获得的聚酯产量最高,分别达到9.12 g/L和7.37 g/L;更高浓度的丁酸抑制了细胞生长,50 g/L丁酸条件下的聚酯产量下降至4.00 g/L以下。此外,Halomonassp.TD08可以利用丁酸和丙酸为混合碳源合成3-羟基丁酸和3-羟基戊酸共聚酯,但丙酸对细胞生长具有较大毒性,2 g/L丙酸和20 g/L丁酸为混合碳源时的细胞干重仅为0.83 g/L,聚酯产量仅为0.15 g/L。通过添加甘油为辅助碳源,明显改善了细胞生长,使得共聚酯的产量提高至3.95 g/L,其中3-羟基戊酸单体含量为8.76 mol%。短链挥发性脂肪酸在盐单胞菌生产聚羟基脂肪酸酯中具有良好的应用前景。     

Utilization of mustard waste isolates for improved production of astaxanthin by Xanthophyllomyces dendrorhous

Astaxanthin production in the wild strain Xanthophyllomyces dendrorhous TISTR 5730 was investigated using different mustard waste media, including mustard waste residue extract (MRE), mustard waste residue hydrolysate (MRH), mustard waste precipitated extract (MPE), and mustard waste precipitated hydrolysate (MPH). The growth of X. dendrorhous and the production of astaxanthin were dependent on the type and initial concentrations of mustard waste media. The MPH medium was the best substrate resulting in yields of biomass and astaxanthin of 19.6 g/L and 25.8 mg/L, respectively, under optimal conditions. MPH medium improved astaxanthin production 11-fold compared to the commonly used commercial yeast malt medium, and 1.3–2.1-fold compared to other mustard waste media.