Protocol to analyze the inhibitory effect of free organic carbon generated by chemoautotrophic bacteria on their CO2 fixation

Saiwei Zhang; Xiaohua Fu; Ya-Nan Wang; Lei Wang

doi:10.1016/j.xpro.2023.102287

Protocol to analyze the inhibitory effect of free organic carbon generated by chemoautotrophic bacteria on their CO₂ fixation

STAR Protoc. 2023 May 5;4(2):102287. doi: 10.1016/j.xpro.2023.102287. Online ahead of print.

Authors

Saiwei Zhang¹, Xiaohua Fu², Ya-Nan Wang³, Lei Wang⁴

Affiliations

¹ Shanghai Academy of Environmental Sciences, Shanghai 200233, China; State Key Laboratory of Pollution Control and Resource Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai 200092, China.
² State Key Laboratory of Pollution Control and Resource Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai 200092, China; Shanghai Institute of Pollution Control and Ecological Security, Shanghai 200092, China.
³ Qingdao Solid Waste Pollution Control and Resource Engineering Research Center, College of Environmental and Municipal Engineering, Qingdao University of Technology, Qingdao 266033, China.
⁴ State Key Laboratory of Pollution Control and Resource Reuse, College of Environmental Science and Engineering, Tongji University, Shanghai 200092, China; Shanghai Institute of Pollution Control and Ecological Security, Shanghai 200092, China. Electronic address: celwang@tongji.edu.cn.

Abstract

Here, we present a protocol to analyze the inhibition of self-generated extracellular free organic carbon (EFOC) on CO₂ fixation by chemoautotrophic bacteria. We detail the construction and operation of membrane reactor, followed by a simulation experiment to verify the inhibition of EFOC on CO₂ fixation. We further describe the analysis of main inhibitory components in EFOC and measurement of abundance and transcription level of ribulose bisphosphate carboxylase/oxygenase (RuBisCO) gene to clarify the mechanism of the main inhibition components on CO₂ fixation. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).¹.

Keywords: Biotechnology and bioengineering; Environmental sciences; Gene Expression; Mass Spectrometry; Microbiology; Molecular Biology.