Bacillus sp. Protein Production via yitMOP and sdpABC Operon Deletion
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Solution Overview
Problem
Current methods for optimizing protein production in Bacillus sp. cells, such as Bacillus subtilis, do not adequately enhance protein yields, limiting industrial biotechnology applications where increased protein production is crucial.
Innovation Solution
Genetically modifying Bacillus sp. cells by introducing modifications to the yitMOP operon, sdpABC operon, or yitM gene to increase protein production, including deletions and disruptions that enhance protein productivity, such as combining deletions of the yitMOP and sdpABC operons or disrupting the yitM gene, which results in increased production of proteins like proteases and amylases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If current optimization methods are used for protein production in Bacillus sp. cells, then the production process is simple, but protein yields are insufficient
Solution Approach 1:
The patent applies the 'Taking out' principle by removing specific genes (yitM, sdpA, sdpB, sdpC) from the Bacillus sp. genome that are responsible for producing unwanted substances. By extracting and eliminating these specific genetic elements, the patent achieves enhanced protein yield without adding complex foreign genetic material, thus resolving the contradiction between increasing protein quantity and maintaining system simplicity
Solution Approach 2:
The patent applies 'Parameter changes' by modifying the genetic composition parameters of the Bacillus sp. cells. Specifically, it changes the presence/absence parameters of certain genes (yitM, sdpABC operon) to alter the cellular metabolism and resource allocation, thereby increasing protein yield. This approach achieves quantitative improvement through qualitative genetic parameter changes
2Quantity of substance
If genetic modifications are introduced to enhance protein production, then protein yield increases, but the risk of unwanted side effects increases
Solution Approach 1:
The patent applies 'Blessing in disguise' by converting the harmful effect of the yitM and sdpABC genes (which produce toxic by-products and compete for resources) into a benefit. By deleting these genes, the patent eliminates the production of harmful substances and redirects the cellular resources toward productive protein synthesis, thus converting a harmful genetic element into a beneficial outcome for protein production
Data Source
AI summary
The present disclosure is generally related to compositions and methods for constructing and obtaining Bacillus sp. host cells (strains) having enhanced protein production phenotypes. Certain embodiments of the disclosure are therefore related to genetically modified Bacillus sp. cells derived (obtained) from parental Bacillus sp. cells (e.g., such as parental Bacillus cells expressing/producing a protein of interest). Thus, certain embodiments are directed to modified Bacillus sp. cells expressing/producing an endogenous protein of interest, or a heterologous protein of interest, wherein the modified Bacillus sp. cell comprises a genetic modification of a yitMOP operon, a combined genetic modification of a yitMOP operon and a sdpABC operon, a genetic modification of a yitM gene and the like, wherein the modified Bacillus sp. cells produce an increased amount of the POI relative the parental cells from which they were derived (i.e., when grown/cultivated/fermented under identical conditions).