Work

Engineering Escherichia coli for controlled autolytic release of intracellular proteins: bacteriophage lambda SRRz versus the 2PaT4L chimeolysin

Year: 2026

Type: article

Abstract: Programmed cell autolysis can simplify the recovery of intracellular recombinant proteins, but differences in experimental conditions and evaluation criteria often limit direct comparisons of lysis systems. This study presents a multicriteria comparison of the two distinct lysis genes, the phage lambda SRRz cassette and the engineered 2PaT4L chimeolysin under identical IPTG-inducible conditions in Escherichia coli . The 2PaT4L outperformed the SRRz across multiple metrics, including cell viability, turbidity, protein release, and escape frequency. Using the inverse of escape frequency as a quantitative measure of lysis efficiency, 2PaT4L and SRRz reached (99.99 ± 0.012)% and (71.10 ± 7.370)% lysis, respectively. Protein release into the culture medium was also higher for 2PaT4L , reaching 1.62 mg/mL after 24 h compared with 1.35 mg/mL for SRRz . These results identify 2PaT4L as the more effective autolysis system under the conditions tested and highlight its potential for intracellular protein recovery. Techno-economic analysis and sensitivity analyses of lysis efficiency, autolysis duration, and inducer concentration indicated that optimizing these conditions can significantly reduce enzyme production costs.

Source: Systems Microbiology and Biomanufacturing

DOI: 10.1007/s43393-026-00550-9

Authors: Kingstone Nihaka, Gugulethu N. Z. Buthelezi, Marilize Le Roes-Hill, Tukayi Kudanga

Institutions: Durban University of Technology, Cape Peninsula University of Technology

Language: English

Topic: Bacterial Genetics and Biotechnology

Subfield: Genetics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,290

Funders: Durban University of Technology, National Research Foundation