Enhancement of monacolin K production via intergeneric protoplast fusion between Aspergillus terreus and Monascus anka

Intergenric protoplast fusion between Aspergillus terreus CA99 and Monascus anka M-3, the high and low producers of monacolin K respectively，was performed for enhancement of monacolin K production. The 24-hour-old mycelia of A. terreus CA99 and M. anka M-3 were treated with 0.5% lywallzyme, 0.3% snailase and 0.3% cellulase at 34℃ for 5 h and at 30℃ for 3.5 h, and their protoplasts formation reached 1.76×10⁷/mL and 1.68×10⁷/mL respectively. Parental protoplasts were irradiated with a 30 W UV light away from 30 cm for 3 min and then mixed. The mixture was incubated with 30% PEG 6000 for 15 min. The reviving fusants were isolated on the regeneration plates. Of the 363 fusants isolated, over 100 showed enhanced monacolin K production compared with the parental strain M. anka M-3. Ten of them produced monacolin K about 1.6-fold of that M.anka M-3 does and the monacolin K titer of two fusants (F49 and F104) increased by about 1-fold. The monacolin K yields of F49 and F104 were 460 μg/mL and 457 μg/mL respectively. In optimized fermentation medium, the monacolin K titer of F49 reached 1216 μg/mL.     

Enhancement of monacolin K production via intergeneric protoplast fusion between Aspergillus terreus and Monascus anka

Intergenric protoplast fusion between Aspergillus terreus CA99 and Monascus anka M-3, the high and low producers of monacolin K respectively,was performed for enhancement of monacolin K production. The 24-hour-old mycelia of A. terreus CA99 and M. anka M-3 were treated with 0.5% lywallzyme, 0.3% snailase and 0.3% cellulase at 34℃ for 5 h and at 30℃ for 3.5 h, and their protoplasts formation reached 1.76×107/mL and 1.68×107/mL respectively. Parental protoplasts were irradiated with a 30 W UV light away from 30 cm for 3 min and then mixed. The mixture was incubated with 30% PEG 6000 for 15 min. The reviving fusants were isolated on the regeneration plates. Of the 363 fusants isolated, over 100 showed enhanced monacolin K production compared with the parental strain M. anka M-3. Ten of them produced monacolin K about 1.6-fold of that M.anka M-3 does and the monacolin K titer of two fusants (F49 and F104) increased by about 1-fold. The monacolin K yields of F49 and F104 were 460 μg/mL and 457 μg/mL respectively. In optimized fermentation medium, the monacolin K titer of F49 reached 1216 μg/mL.     

链霉菌种间原生质体融合的研究(Ⅱ)——融合子56-2菌株的鉴定

通过金霉素链霉菌和龟裂链霉菌原生质体融合选育出的融合子５６－２菌株，其酯酶同工酶谱与双亲有显著差异，产抗生素能力及遗传稳定性均优于金霉素链霉菌３＃，发酵产物四环素符合中国药典的各项指标，并具有抗噬菌体Ｐ５，Ｐ８的能力。     

SD－5菌株原生质体形成与再生条件的研究

探讨了培养基成分、培养时间、高渗缓冲液成分和ｐＨ、溶菌酶浓度以及酶解温度和时间对苏云金杆菌ＳＤ—５菌株原生质体形成和再生的影响；结果表明，ＳＤ—５菌株原生质体形成和再生的最佳条件组合为：ＰＢＧ培养基、３０℃培养１４ｈ再活化５ｈ，以ＳＭＭｐＨ６．５作酶解缓冲液，溶菌酶浓度０．６ｍｇ／ｍｌ，３４℃处理６０ｍｉｎ．     

Rhizoctonia solani Kuhn原生质体的制备和再生

应用市售商品酶,从立枯丝核菌(Rhizoctonia solani)菌丝体中,成功地分离出高产的原生质体。日本产Cellulase “onozuka”R—101%与上海产蜗牛酶1～1.5%或者与上海产溶菌酶1～1.5%组合,在35℃,0.6M MgSO_4·7H_2O条件下,菌丝酶解效果很好。菌龄,酶及酶解条件以及培养基的成分等因素与原生质体的形成及再生密切相关。对原生质体的形成过程及再生过程的形态学变化进行了观察。二种原生质体再生转化形式,在再生培养基中能够观察得到。     

细菌科间原生质体融合方法的研究

本文通过对枯草芽孢杆菌(Bacillus subtilis)151衍生株和北京棒杆菌(Corynebacterium pekinese)1134衍生株的科间原生质体融合的研究以及灭活原生质体的科间融合的研究,揭示了细菌科间原生质体融合的可能性,同时进行了不同融合方法的分析对比,为今后工业微生物的远缘育种提供了可以选择的有效途径和理论依据。     

建筑形象的原型特征

本讨论了建筑形式的原型问题．这对建筑的设计和创作有重要的作用。根据W·冯特的构造心理学，感情可以分为三度。即激动或沉静，紧张或松弛．愉快或不谕快。根据感情三度原理．建筑形式也可分为三种形态来分析，即高度方向．横向和纵向。     

G-8菌株原生质体形成和再生条件的研究

以浮游球衣细菌Ｇ－８菌为出发菌株,有杉甘氨酸－溶菌酶－ＥＤＴＡ方法,研究了各种因素对原生质体形成和再生的影响。在原生质体形成和再生的最佳条件下制备原生质体,Ｇ－８菌原生质体形成率和一率分别达９６．８％和２９．８％,通过光学显微镜技术,观察了原生质体形成和再生的过程。