Key Laboratory of Systems Bioengineering, Ministry of Education (Tianjin University), Tianjin, 300072, China.
School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.
Sci Rep. 2017 Jul 21;7(1):6189. doi: 10.1038/s41598-017-06537-w.
Nisin, one kind of natural antimicrobial peptide, is produced by certain Lactococcus lactis strains, which generally require expensive high-quality nitrogen sources due to limited ability of amino acids biosynthesis. Here we use defatted soybean meal (DSM) as sole nitrogen source to support L. lactis growth and nisin production. DSM medium composition and fermentation conditions were optimized using the methods of Plackett-Burman design and central composite design. The highest nisin production of 3879.58 IU/ml was obtained in DSM medium, which was 21.3% higher than that of commercial medium. To further increase the utilization ability of nitrogen sources, we enhanced the proteolytic function in L. lactis through rationally expressing the related enzymes, which were selected according to the compositions of amino acids and molecular weight of peptides in DSM medium. Significantly, an artificial proteolytic system consisting of a heterologous protease (NprB), an oligopeptides transporter subunit (OppA) and two peptidases (PepF and PepM) was introduced into L.lactis. The constructed strain BAFM was capable of achieving efficient biomass accumulation and nisin yield with 30% decreased amount of DSM hydrolysates, which further reduced the cost of nisin production. The strategy described here offers opportunities for low-cost L. lactis fermentation and large-scale nisin production in industry.
乳链菌肽(Nisin)是由某些乳球菌菌株产生的一种天然抗菌肽,由于其氨基酸生物合成能力有限,通常需要昂贵的高质量氮源。在这里,我们使用脱脂豆粕(DSM)作为唯一的氮源来支持乳链菌的生长和乳链菌肽的生产。我们使用 Plackett-Burman 设计和中心组合设计方法优化了 DSM 培养基的组成和发酵条件。在 DSM 培养基中获得了最高的 3879.58 IU/ml 的乳链菌肽产量,比商业培养基高 21.3%。为了进一步提高氮源的利用能力,我们通过合理表达相关酶来增强乳链菌的蛋白水解功能,这些酶是根据 DSM 培养基中氨基酸的组成和肽的分子量选择的。值得注意的是,我们将一个由异源蛋白酶(NprB)、寡肽转运亚基(OppA)和两种肽酶(PepF 和 PepM)组成的人工蛋白水解系统引入到乳链菌中。构建的 BAFM 菌株能够实现高效的生物量积累和乳链菌肽的产量,DSM 水解物的用量减少了 30%,进一步降低了乳链菌肽的生产成本。这里描述的策略为低成本的乳链菌发酵和工业大规模生产乳链菌肽提供了机会。