Song Xiaofen, Wei Hansheng, Zhou Yueyue, Song Weiwei, Shi Ce, Mu Changkao, Wang Chunlin, Wang Xiaopeng
Marine Economic Research Center, Donghai Academy, Ningbo University, No. 169, Qixing South Road, Meishan Port District, Beilun District, Ningbo 315000, China.
Key Laboratory of Aquacultral Biotechnology, Chinese Ministry of Education, Ningbo University, No. 169, Qixing South Road, Meishan Port District, Beilun District, Ningbo 315000, China.
Int J Mol Sci. 2025 Mar 12;26(6):2543. doi: 10.3390/ijms26062543.
With the development of the aquatic products processing industry, 6-8 million tons of shrimp and crab shell waste are produced globally annually, but, due to the lack of high-value conversion technology, crab shells are often discarded in large quantities as a by-product of processing. -based microbial cell factories are capable of biosynthesis of high-value products using a wide range of substrates; however, there is currently no reliable fermentation model for producing high-value chemicals using crab shell waste by strains. In this study, we first explored the culture conditions of shell fermentation using KT2440 through single-factor and orthogonal experiments, and the optimized fermentation parameters obtained are given as follows: a temperature of 30 °C, fermentation time of 42 h, substrate solid-liquid ratio of 7%, and rotational speed of 200 rpm. After optimization, the maximum cell growth was increased by 64.39% from 350.67 × 10 CFU/mL to 576.44 × 10 CFU/mL. Combined with engineering modification, two engineered strains, KT and KT, expressing exogenous proteases, were obtained, and the maximum growth was increased from 316.44 × 10 CFU/mL to 1268.44 × 10 CFU/mL and 616.89 × 10 CFU/mL, which were 300.84% and 94.94% higher, respectively. In addition, the engineered strain KT, which regulates nitrogen metabolism, was obtained, and the accumulation of intracellular polyhydroxy fatty acid esters (PHA) was increased from 20.00 mg/L to 78.58 mg/L, which was a significant increase of 292.93% relative to the control group. This study provides a theoretical basis and technical support for the high-value utilization of shrimp and crab shell resources and the development of environmentally friendly bioproducts.
随着水产品加工业的发展,全球每年产生600 - 800万吨虾蟹壳废弃物,但由于缺乏高值转化技术,蟹壳常作为加工副产品被大量丢弃。基于微生物的细胞工厂能够利用多种底物生物合成高值产品;然而,目前尚无可靠的利用蟹壳废弃物通过菌株生产高值化学品的发酵模型。在本研究中,我们首先通过单因素和正交实验探索了利用KT2440进行壳发酵的培养条件,得到的优化发酵参数如下:温度30℃,发酵时间42小时,底物固液比7%,转速200转/分钟。优化后,最大细胞生长量从350.67×10 CFU/mL提高到576.44×10 CFU/mL,增加了64.39%。结合工程改造,获得了两种表达外源蛋白酶的工程菌株KT和KT,最大生长量分别从316.44×10 CFU/mL提高到1268.44×10 CFU/mL和616.89×10 CFU/mL,分别提高了300.84%和94.94%。此外,获得了调节氮代谢的工程菌株KT,细胞内聚羟基脂肪酸酯(PHA)的积累从20.00 mg/L增加到78.58 mg/L,相对于对照组显著增加了292.93%。本研究为虾蟹壳资源的高值利用和环保型生物产品的开发提供了理论依据和技术支持。