Increased astaxanthin production by a Phaffia rhodozyma mutant grown on date juice from Yucca fillifera

The wild strain and the astaxanthin-overproducing mutant strain 25–2 of Phaffia rhodozyma were analyzed in order to assess their ability to grow and synthesize astaxanthin in a minimal medium composed of g L⁻¹: KH₂PO₄ 2.0; MgSO₄ 0.5; CaCl₂ 0.1; urea 1.0 and supplemented with date juice of Yucca fillifera as a carbon source (yuca medium). The highest astaxanthin production (6170 μg L⁻¹) was obtained at 22.5 g L⁻¹ of reducing sugars. The addition of yeast extract to the yuca medium at concentrations of 0.5–3.0 g L⁻¹ inhibited astaxanthin synthesis. The yuca medium supported a higher production of astaxanthin, 2.5-fold more than that observed in the YM medium. Journal of Industrial Microbiology & Biotechnology (2000) 24, 187–190. Received 14 July 1999/ Accepted in revised form 02 December 1999     

60Coγ射线诱变选育高产虾青素红发夫酵母突变株

为了筛选高产虾青素红发夫酵母突变株,用不同剂量^60Coγ射线对出发菌株菌液进行反复辐射处理,得到突变株W6-318,并对其生物学特性进行了研究。结果表明不同照射剂量下正突变率为10％—36％,在射线诱变剂量为3．5kGy时诱变效果最佳。最优化条件下突变株生物量、总类胡萝卜素和虾青素产量分别为10．15gL^-1、14．97mgL^-1和12．55mgL^-1,分别比出发菌株提高11．17％、86．39％和101．8％。5L发酵罐放大培养中的生物量、总类胡萝卜素和虾青素产量分别为15．56gL^-1、18．54mgL^-1和14．97mgL^-1。     

红发夫酵母积累虾青素的代谢调控机理研究进展

由红发夫酵母合成的虾青素是一种非常有商业价值的类胡萝卜素.综述了近年来国内外在红发夫酵母中虾青素的生物合成途径、合成代谢调控机理等领域的相关研究进展,并提出了国内应开展的相关创新性研究.     

生物技术生产虾青素研究的最新进展

虾青素是一种具有极强的生物抗氧化性的酮式类胡萝卜素在医药,食品,化妆品等方面有着极广阔的应用前景。对目前生物技术生产虾青素的两种主要方法：即雨生红球藻(haematococcus pluvialis)培养法和红发夫酵母(xanthopltyllomyces dendrorhous,formedy Phaffia raodazyma)发酵法的最新研究进展,包括采用的微生物的生理代谢特性、虾青素合成途径、生产技术特点、工程菌种的构建等方面进行了系统的总结。针对天然虾青素生产技术研究中存在的问题和不足,提出了建议和解决的方法。     

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。     

Simultaneous Determination of Diphenyl and o-Phenylphenol in Citrus Fruits

A simple method for simultaneous determination of diphenyl and o-phenyl phenol in citrus fruits was established. Fruits were distilled with a distillable oil analyzer. The citrus fruit extract was taken from this apparatus and anthracene was added as an internal standard. Gas chromatography was carried out with a column packed with 10% FFAP and a flame ionization detector. Column temperature was increased from 150 to 210°C. The retention times of diphenyl, o-phenylphenol and anthracene were 4.2, 14.0 and 15.4 min, respectively.

This method was completed within 2.5 hr and applied to samples of grapefruits, lemons, oranges, navel oranges, ponkan, iyokan, hassaku, unshu mikan and amanatsu mikan. In the recovery tests with these fruits, diphenyl was recovered in the range from 90.1 to 96.9% and o-phenylphenol was recovered in the range from 86.5 to 99.3%.     

法夫酵母高产虾青素的选育及发酵条件的优化

对法夫酵母原始菌株进行了紫外线－氯化锂复合诱变,获得了一株遗传性状稳定、在25℃条件下虾青素高产的菌株UL-40。其类胡萝卜素产量为7.10mg/L,经HPLC测定,其虾青素产量为643.97μg/g,比出发菌株提高了249.87%。利用SAS软件中的Plackett-Burman设计对发酵温度、初始pH值以及发酵培养基的八种组分进行优化组合,从中选出发酵温度、初始pH值和Corn steep liquor浓度为重要因素,通过响应面分析法建立了模型并从该模型中计算出在发酵温度为16.78℃、初始pH为4.73和CSL浓度为7.06mg/L时预测的最大响应值为3.9407mg/L,经实验证实此点的实测值为3.9261mg/L,证明模型是有效的并且存在极大值点。采用优化后的发酵条件及发酵培养基,法夫酵母生产虾青素的产量较原始发酵培养基提高了20.4%。     

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。     

Improvement of astaxanthin production by a newly isolated Phaffia rhodozyma mutant with low-energy ion beam implantation

Aims: Isolation, characterization and identification of Phaffia sp. ZJB 00010, and improvement of astaxanthin production with low‐energy ion beam implantation. Methods and Results: A strain of ZJB 00010, capable of producing astaxanthin, was isolated and identified as Phaffia rhodozyma, based on its physiological and biochemical characteristics as well as its internal transcribed spacer (ITS) rDNA gene sequence analysis. With low‐energy ion beam implantation, this wild‐type strain was bred for improving the yield of astaxanthin. After ion beam implantation, the best mutant, E5042, was obtained. The production of astaxanthin in E5042 was 2512 μg g^?1 (dry cell weight, DCW), while the wild‐type strain was about 1114 μg g^?1 (DCW), an increase of 125·5%. Moreover, the fermentation conditions of mutant E5042 for producing astaxanthin were optimized. The astaxanthin production under the optimized conditions was upscaled and studied in a 50‐l fermentor. Conclusions: A genetically stable mutant strain with high yield of astaxanthin was obtained using low‐energy ion beam implantation. This mutant may be a suitable candidate for the industrial‐scale production of astaxanthin. Significance and Impact of the Study: Astaxanthin production in Phaffia rhodozyma could be fficiently improved by low‐energy ion beam implantation, which is a new technology in the mutant breeding of micro‐organisms. The mutant obtained in this work could potentially be utilized in industrial production of astaxanthin.