Division of Environmental Science & Ecological Engineering, Korea University, Seoul 02841, Republic of Korea.
Department of Biomedical Sciences, Korea University, Seoul 02841, Republic of Korea.
Bioresour Technol. 2021 Jun;330:124974. doi: 10.1016/j.biortech.2021.124974. Epub 2021 Mar 17.
Genetic manipulation of the Porphyridium sp. may increase the production of phycoerythrin. Since phycobiliproteins capture and transfer energy to both photosystems (PS I and PS II), it was hypothesized that the gene mutation involved increases phycoerythrin synthesis. The gene encoding chlorophyll synthase (CHS1) was selected as chlorophyll synthase plays an important role in photosynthesis, mediating the final process of chlorophyll synthesis. Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 ribonucleoprotein (CRISPR/Cas9 RNP) delivery system was used to generate the chlorophyll synthase loss-of-function mutants (Δchs1). Independent Δchs1 showed no differences in the growth and production of sulfated polysaccharide compared to control. Phycoerythrin contents of the two independent mutants substantially increased regardless of light source. This study provides a novel applicability for the CRISPR/Cas9 RNP method in red microalgae toward a bio-product of interest. The obtained mutants could serve as potential producers of phycoerythrin if Porphyridium is selected as a natural source.
对紫球藻进行基因操作可能会增加藻红蛋白的产量。由于藻胆蛋白可以同时捕获和传递能量给两个光系统（PS I 和 PS II），因此推测所涉及的基因突变会增加藻红蛋白的合成。选择编码叶绿素合酶（CHS1）的基因为研究对象，是因为叶绿素合酶在光合作用中起着重要作用，介导了叶绿素合成的最后过程。成簇规律间隔短回文重复序列/CRISPR 相关蛋白 9 核糖核蛋白（CRISPR/Cas9 RNP）递送系统被用来产生叶绿素合酶功能丧失突变体（Δchs1）。与对照相比，独立的Δchs1在生长和硫酸多糖的产生方面没有差异。无论光源如何，两个独立突变体的藻红蛋白含量都大大增加。本研究为 CRISPR/Cas9 RNP 方法在红色微藻中作为生物制品的应用提供了新的可能性。如果选择紫球藻作为天然来源，获得的突变体可以作为藻红蛋白的潜在生产者。
