<i>Spirulina</i>-in Silico-Mutations and Their Comparative Analyses in the Metabolomics Scale by Using Proteome-Based Flux Balance Analysis

• Main Authors: Supatcha Lertampaiporn, Jittisak Senachak, Wassana Taenkaew, Chiraphan Khannapho, Apiradee Hongsthong
• Format: Article
• Language: English
• Published: MDPI AG 2020-09-01
• Series: Cells
• Subjects: genome-scale; flux balance analysis; proteome analysis; temperature response; histidine kinase; in silico mutation
• Online Access: https://www.mdpi.com/2073-4409/9/9/2097

This study used an in silico metabolic engineering strategy for modifying the metabolic capabilities of <i>Spirulina</i> under specific conditions as an approach to modifying culture conditions in order to generate the intended outputs. In metabolic models, the basic metabolic fluxes in steady-state metabolic networks have generally been controlled by stoichiometric reactions; however, this approach does not consider the regulatory mechanism of the proteins responsible for the metabolic reactions. The protein regulatory network plays a critical role in the response to stresses, including environmental stress, encountered by an organism. Thus, the integration of the response mechanism of <i>Spirulina</i> to growth temperature stresses was investigated via simulation of a proteome-based GSMM, in which the boundaries were established by using protein expression levels obtained from quantitative proteomic analysis. The proteome-based flux balance analysis (FBA) under an optimal growth temperature (35 °C), a low growth temperature (22 °C) and a high growth temperature (40 °C) showed biomass yields that closely fit the experimental data obtained in previous research. Moreover, the response mechanism was analyzed by the integration of the proteome and protein–protein interaction (PPI) network, and those data were used to support in silico knockout/overexpression of selected proteins involved in the PPI network. The <i>Spirulina</i>, wild-type, proteome fluxes under different growth temperatures and those of mutants were compared, and the proteins/enzymes catalyzing the different flux levels were mapped onto their designated pathways for biological interpretation.