Genetically Modified Phage for Stable Biomolecule Production
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Solution Overview
Problem
Existing methods for producing biomolecules using bacteriophage systems often result in phage contamination and bacterial lysis, which complicates pharmaceutical applications, requires shutdown of production lines, and reduces recombinant protein yield.
Innovation Solution
A genetically modified phage with deleted Int, S, R, Rz, Xis genes, and optionally Q gene, integrated into the bacterial host cell genome, to prevent lytic properties and contamination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If a lambda DE3 lysogen phage is integrated in the bacterial host cell to enable T7 RNA polymerase expression, then the expression of target protein is enabled, but the phage may recover its lytic properties causing bacterial lysis and phage contamination
Solution Approach 1:
The patent removes specific genes (Int, S, R, Rz, Xis, and optionally Q) from the lambda phage genome to eliminate the lytic cycle capability while preserving the ability to maintain the T7 RNA polymerase gene in the host. This extraction of harmful genetic elements resolves the contradiction by preventing phage recovery of lytic properties while maintaining expression functionality.
Solution Approach 2:
The phage genome is segmented into essential functional regions (maintained) and harmful lytic regions (removed). The patent creates a modified phage construct where only the necessary portions for maintaining T7 RNA polymerase expression are retained, while the segments responsible for lytic cycle (Int, S, R, Rz, Xis, Q genes) are deleted, thereby achieving stable lysogeny without lysis risk.
2Reliability
If bacterial lysis occurs during protein production, then the culture must be shutdown and production lines renewed, but this reduces productivity and increases production time
Solution Approach 1:
The patent performs preliminary genetic modification of the phage by deleting lytic genes before the phage integrates into the host genome. This preliminary action prevents the occurrence of bacterial lysis during protein production, thereby avoiding shutdowns and maintaining continuous productivity without compromising culture stability.
3Object-affected harmful factors
If decontamination process is performed to remove phage contamination, then product safety is improved, but this requires shutdown of production lines and complete renewal of culture batches
Solution Approach 1:
The patent applies preliminary anti-action by genetically engineering the phage to eliminate its lytic capability before it can cause contamination. By deleting the Int, S, R, Rz, Xis, and Q genes, the phage is rendered incapable of initiating the lytic cycle and producing contamination, thus preventing the need for decontamination procedures and associated production downtime.
4Reliability
If modified lambda phages with deleted genes are used, then some lytic properties are reduced, but lysis and contamination may still occur
Solution Approach 1:
The patent applies parameter changes by systematically removing multiple critical genes (Int, S, R, Rz, Xis, and optionally Q) from the phage genome. This multi-gene deletion strategy fundamentally changes the phage's biological parameters, completely eliminating its ability to undergo lytic cycle and cause contamination, thereby achieving absolute lytic control and prevention of harmful effects.
Data Source
Figure 1
AI summary
The present invention relates to a genetically modified phage and use thereof in a method for producing a biomolecule of interest.