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Other Glycolytic Pathways

Other Glycolytic Pathways

The pentose phosphate pathway (PPP) operates in parallel with glycolysis, facilitating the metabolism of both pentoses and glucose. This pathway consists of two distinct phases: the oxidative and non-oxidative phases. While it does not directly generate ATP, the intermediates formed during the process can integrate into glycolysis, contributing to cellular energy metabolism when required.Oxidative Phase: NADPH ProductionThe oxidative phase of the pentose phosphate pathway is primarily...

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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

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Adaptive laboratory evolution optimizes an engineered phosphite utilization pathway in Synechococcus elongatus PCC

Hiroki Murakami¹, Naoki Momokawa², Kei Motomura²

¹Unit of Biotechnology, Graduate School of Integrated Sciences for Life, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8530, Japan; Faculty of Global Interdisciplinary Science and Innovation, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.

Journal of Bioscience and Bioengineering

|December 13, 2025

Summary

Long-term cultivation improved a phosphite-dependent microbial strain, enhancing its growth and phosphite consumption. Mutations in transporter genes boosted phosphite uptake without affecting phosphate specificity, optimizing biocontainment.

Keywords:

Adaptive laboratory evolution Biocontainment Cyanobacteria Phosphite transporter

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Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

Area of Science:

Synthetic biology
Microbial genetics
Metabolic engineering

Background:

Metabolic engineering in synthetic biology can cause burdens, leading to adaptive evolution.
A phosphite (Pt)-dependent system was developed for microbial biocontainment but caused growth retardation.

Purpose of the Study:

Investigate the effects of long-term cultivation on a Pt-dependent Synechococcus elongatus strain.
Identify genetic adaptations that improve growth and Pt utilization.

Main Methods:

Serial passaging of the Pt-dependent strain RH714.
Comparative growth rate and Pt consumption analysis.
Sequence analysis of transporter genes (htxBCDE) and genetic reintroduction.

Main Results:

Passaged RH714 showed improved growth and higher Pt consumption compared to the original strain.
Point mutations in htxBCDE genes were identified as responsible for enhanced Pt transport.
Reintroduction of mutated genes into wild-type S. elongatus PCC 7942 replicated the improved phenotype.

Conclusions:

Long-term passage cultivation selects for optimized mutants with enhanced Pt metabolism.
Mutations in htxBCDE genes improve Pt transport activity without altering phosphate (Pi) specificity.
Developed strains maintain biocontainment capabilities while exhibiting improved proliferation.