Montiel-González Alba Mónica, Fernández Francisco José, Viniegra-González Gustavo, Loera Octavio
Department of Biotechnology, Universidad Autónoma Metropolitana-Iztapalapa, Mexico DF, México.
Appl Biochem Biotechnol. 2002 Jul-Dec;102-103(1-6):63-70. doi: 10.1385/abab:102-103:1-6:063.
Invertase production by Aspergillus niger grown by solid-state fermentation was found to be higher than by conventional submerged fermentation. The haploid mutant strains Aw96-3 and Aw96-4 showed better productivity of various enzymes, as compared to wild-type parental strain A. niger C28B25. Here we use parasexual crosses of those mutants to increase further the productivity of invertase in solid-state fermentation. We isolated both a diploid (DAR2) and an autodiploid (AD96-4) strain, which were able to grow in minimal medium after mutation complementation of previously isolated haploid auxotrophic strains. Invertase production was measured in solid-state fermentation cultures, using polyurethane foam as an inert support for fungal growth. Water activity value (Aw) was adjusted to 0.96, since low Aw values are characteristic in some solid-state fermentation processes. Such diploid strains showed invertase productivity levels 5-18 times higher than levels achieved by the corresponding haploid strains. For instance, values for C28B25, Aw96-3, Aw96-4, DAR2, and AD96-4 were 441, 254, 62, 1324, and 2677 IU/(L x h), respectively. These results showed that genetic recombination, achieved through parasexual crosses in A. niger, results in improved strains with potential applications for solid-state fermentation processes.
通过固态发酵培养的黑曲霉产生的转化酶比传统的深层发酵更高。与野生型亲本菌株黑曲霉C28B25相比,单倍体突变菌株Aw96 - 3和Aw96 - 4表现出更好的各种酶的生产能力。在这里,我们利用这些突变体的准性杂交来进一步提高固态发酵中转化酶的生产能力。我们分离出了一个二倍体(DAR2)和一个同源二倍体(AD96 - 4)菌株,它们在对先前分离的单倍体营养缺陷型菌株进行突变互补后能够在基本培养基中生长。在固态发酵培养物中测量转化酶的产量,使用聚氨酯泡沫作为真菌生长的惰性支持物。将水分活度值(Aw)调整到0.96,因为在一些固态发酵过程中低Aw值是其特征。这样的二倍体菌株显示出的转化酶生产能力水平比相应单倍体菌株所达到的水平高5 - 18倍。例如,C28B25、Aw96 - 3、Aw96 - 4、DAR2和AD96 - 4的值分别为441、254、62、1324和2677 IU/(L×h)。这些结果表明,通过黑曲霉中的准性杂交实现的基因重组产生了具有固态发酵过程潜在应用价值的改良菌株。