Mehta Bina J, Obraztsova Irina N, Cerdá-Olmedo Enrique
Departamento de Genética, Universidad de Sevilla, E-41080 Seville, Spain.
Appl Environ Microbiol. 2003 Jul;69(7):4043-8. doi: 10.1128/AEM.69.7.4043-4048.2003.
The industrial production of beta-carotene with the zygomycete Blakeslea trispora involves the joint cultivation of mycelia of opposite sex in the presence of beta-ionone and other chemical activators. We have obtained improved strains by mutation and heterokaryosis. We chose wild strains on the basis of their growth and carotene content in single and mated cultures. Following exposure of their spores to N-methyl-N'-nitro-N-nitrosoguanidine, we obtained high-carotene mutants, which were more productive than their parents but similar to them in having beta-carotene as the main product. Further increases in carotene content were obtained after a new round of mutagenesis in one of the mutants. The production was shifted to lycopene in cultures incubated in the presence of nicotine and in lycopene-rich mutants derived from the wild strains. The highest production levels were achieved in intersexual heterokaryons, which contained mutant nuclei of opposite sex. These contained up to 39 mg of beta-carotene or 15 mg of lycopene per g (dry mass) under standard laboratory conditions in which the original wild strains contained about 0.3 mg of beta-carotene per g (dry mass). Beta-ionone did not increase the carotene content of these strains. Not all wild strains lent themselves to these improvements, either because they produced few mutants or because they did not increase their carotene production in mated cultures.
利用三孢布拉氏霉菌工业生产β-胡萝卜素涉及在β-紫罗酮和其他化学激活剂存在的情况下对异性菌丝体进行联合培养。我们通过诱变和异核化获得了改良菌株。我们根据野生菌株在单培养和交配培养中的生长情况和胡萝卜素含量来选择它们。将其孢子暴露于N-甲基-N'-硝基-N-亚硝基胍后,我们获得了高胡萝卜素突变体,这些突变体比其亲本产量更高,但与亲本一样以β-胡萝卜素为主要产物。在其中一个突变体中进行新一轮诱变后,胡萝卜素含量进一步提高。在尼古丁存在下培养的培养物以及源自野生菌株的富含番茄红素的突变体中,产物转变为番茄红素。在含有异性突变核的两性异核体中实现了最高产量水平。在标准实验室条件下,这些异核体每克(干重)含有高达39毫克的β-胡萝卜素或15毫克的番茄红素,而原始野生菌株每克(干重)约含有0.3毫克的β-胡萝卜素。β-紫罗酮并未增加这些菌株的胡萝卜素含量。并非所有野生菌株都适合这些改良,要么是因为它们产生的突变体很少,要么是因为它们在交配培养中没有增加胡萝卜素产量。