BE-14106 Biosynthetic Gene Cluster Cloning
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Solution Overview
Problem
The existing technologies face challenges in identifying and cloning the correct gene cluster for the biosynthesis of the macrolactam antibiotic BE-14106, which limits the repertoire of available compounds and their properties, such as efficacy and toxicity, and requires complex sequence analysis and probe design.
Innovation Solution
The cloning and sequencing of the novel BE-14106 biosynthetic gene cluster from a newly isolated Streptomyces strain MP28-13, which includes 22 genes encoding proteins involved in the biosynthesis, allowing for genetic manipulation to modify expression levels and produce novel compounds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If complex sequence analysis and probe design are used to identify and clone the correct gene cluster, then the biosynthesis of macrolactam antibiotics can be achieved, but the process becomes overly complicated and time-consuming
Solution Approach 1:
The patent segments the polyketide synthase gene cluster into distinct modular units, each responsible for specific biosynthetic functions. This segmentation allows for easier identification and manipulation of individual gene modules, simplifying the overall cloning process while maintaining high identification accuracy through systematic analysis of discrete functional elements.
Solution Approach 2:
The patent performs preliminary sequencing and characterization of the gene cluster before attempting cloning. By obtaining the complete nucleotide sequence and identifying open reading frames in advance, the researchers prepared targeted probes and primers that greatly simplified the subsequent cloning process, avoiding the need for complex trial-and-error approaches.
2Adaptability or versatility
If the repertoire of available compounds is limited, then the biosynthetic machinery can be simpler, but the ability to produce derivatives with improved efficacy and reduced toxicity is constrained
Solution Approach 1:
The patent demonstrates that the modular polyketide synthase system can produce multiple different macrolactam compounds by varying the combination and configuration of standardized modules. This multi-functionality allows a single biosynthetic machinery to generate diverse compounds with different biological activities, including derivatives with improved efficacy and reduced toxicity, without requiring completely separate biosynthetic pathways for each compound.
Solution Approach 2:
The patent applies local quality by modifying specific domains within individual modules of the polyketide synthase system to alter the properties of produced compounds. By making targeted changes to specific functional regions (such as ketosynthase or acyltransferase domains) rather than redesigning the entire system, the researchers can produce derivatives with improved properties while maintaining the overall simplicity of the modular architecture.
Data Source
AI summary
A nucleic acid molecule comprises a nucleotide sequence: as shown in SEQ ID No. 1, which is the complement of SEQ ID No. 1, which is degenerate with SEQ ID No. 1, or which has at least 85% sequence identity with SEQ ID No. 1, or which is a part of such a sequence. The nucleic acid molecule encodes or is a complementary to a nucleic acid molecule encoding one or more polypeptides, or comprises or is complementary to a nucleic acid molecule comprising one or more genetic elements, having functional activity in the synthesis of a polyketide-based or macrolactam molecule. The nucleic acid molecule may be used to prepare a modified BE-14106 biosynthetic gene cluster for the preparation of a modified BE-14106 molecule.