Strain Improvement Services
BackgroundService ProcedureOur AdvantagesCase StudyFAQs
Background
Overview of Strain Improvement
Microorganisms can generate thousands of valuable products, including proteins, primary and secondary metabolites. The growing demand for biotechnological products against limited metabolic capacity of industrially used microorganisms has led to an increased interest on strain improvement.
The aim of strain improvement is to short fermentation times, enhance metabolite yield, lower toxicity, improve titer and stability, substrate uptake and tolerance of the strains. Strain improvement also serves other goals, such as the elimination of undesirable products or analogs, discovery of new antibiotics, and deciphering of biosynthetic pathways.
Application of Strain Improvement
Microorganisms produce various essential compounds for the industrial, such as small molecules essential for vegetative growth (primary metabolites), and inessential (secondary metabolites), large molecules such as proteins, nucleic acids, carbohydrate polymers and more. Regulatory mechanisms have evolved in microorganisms that enable a strain to avoid an excessive production of metabolites; thus strain improvement programs are required for commercial application. The strain improvement strategy greatly increases fermentation productivity and significantly reduces costs. These modified strains are also used for other goals, such as eliminating unwanted products or analogs, discovering novel structural compounds, and deciphering biosynthetic pathways.
Creative BioMart Microbe strives to serve clients by providing strain improvement service that meet the highest industry and production expectations. Please feel free to contact us for more information.
Fig. 1. The processes for targeted screening of microbial species and the novel isolation technologies. (Jiang, et al., 2024)
Service Procedure
Platforms
ARTP Mutagenesis Breeding Machine
This instrument has a high mutation rate, compact structure, simple operation, high safety and fast mutagenesis speed. A mutagenesis operation (within a few minutes) can obtain a large capacity mutation library, which greatly improves the mutation intensity and mutation library capacity of strains.
Services Details
With rich experience of microbial service in industry, Creative BioMart Microbe has established a complete strain improvement platform. With rich experienced experts, Creative BioMart Microbe can offer you with a panel of comprehensive solutions.
Mutation Service
As the most commonly method, the main mutation method is to use physical, chemical, biological, or combinations of different mutagenesis to modify the target microorganism's genome to produce mutants. The naturally occurring mutation rate in microorganism is 10-8-10-5, which can be increased to 10-6-10-3 after mutagenesis.
Among numerous mutation services, the mutation efficiency of random mutagenesis is very high. Common mutation techniques include UV laser random mutagenesis and ARTP (Atmospheric Room Temperature Plasma) mutagenesis. In addition, random mutagenesis technology combined with high throughput screening can significantly improve the production efficiency of fermentation strains.
Genetic Recombination Service
Genetic recombination allows the exchange of genetic information between two DNA molecules in a precise, specific, and faithful manner, regardless of size. We provide genetic recombination services by transformation, transduction, conjugation, and protoplast fusion. Among which, Red/ET recombination technology is the most common used method for DNA engineering. Red/ET recombination technology is based on homologous recombination involving a λ phage-derived protein pair, Redα/Redβ, and 50 bp homology regions. It can help you realize deletion, insertion, point mutation at any position because the sequence of the homology regions can be chosen freely.
Overexpression Service
To achieve the high expression of target enzyme or metabolites, overexpression of related genes in target microbial strains is necessary in some cases. Creative BioMart Microbe offer custom one stop strain overexpression service involving gene synthesis, vector construction, transformation, strain screening and verification. The diverse options for vectors with diversified strong promoters and selective markers. Moreover, we can help you realize endogenous overexpression of target genes using the CRISPR activation system. Though overexpression may bring genetic instability, structural instability, and metabolic burden to the host, it brings many advantages for application of improved strains.
Gene Editing Service
Gene editing is a group of technologies that allow us change an organism's DNA. A recent one is known as CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9), which can help you get genetic material to be added, removed, or altered at a particular location. It has gained the favor of many scientists because it is faster, cheaper, more accurate, and more efficient than other existing genome editing methods. Creative BioMart Microbe has developed a well gene editing platform, λ Red-CRISPR/Cas9 system, which leverages the efficiency of traditional λ Red recombineering with the ease of CRISPR/Cas for seamless, targeted genome editing.
In addition, our scientists can help you get improved strains with precise editing by base editors (such as cytosine base editors (CBEs), adenine base editors (ABEs) and glycosylase base editors (GBEs)) or prime editing technology. These technologies don't need double-stranded DNA breaks (DSBs). Furthermore, you can get mutational strains by constructing CRISPR mutation library.
Our Advantages
- Over 20,000 strains in microbial library. Such as Bacillus amyloliquefaciens, Escherichia coli, Bacillus subtilis, Trichoderma reesei, Paenibacillus polymyxa, Pseudomonas fluorescens,Bacillus cereus, Methyltrophic bacillus, Trichoderma hartzii, Bacillus adamant, Marine bacillus, Phytophthora dioides, Bacillus brevis, bacillus licheniformis, etc.
- High-throughput screening platform.
- Industrial sized scale-up.
- Rapid development time.
- High success rate.
Case Study
Case Study 1: High yield of pyrroloquinoline quinone was obtained by Methylobacterium extorquens AM1 treated with ARTP.
In order to improve the efficiency of the PQQ production by Methylobacterium extorquens AM1, the strain was treated by ARTP. Positive mutants with changes in PQQ yield were obtained based on a high-throughput screening approach. After ARTP treatment, the positive mutation rate was 31.6%. Furthermore, an excellent positive mutant M. extorquens AM1 (E-F3) was obtained with the yield of 54.0 mg/L PQQ, which was approximately 3 times as much compared with that of the wild-type strain.
Fig. 2. Batch fermentation of M. extorquens AM1 (A) and E-F3 (B). (Li, et al., 2016)
Case Study 2: The activity of glutaminase was improved by using Streptothrix pentaeniensis produced by ARTP.
Streptomyces spore was mutated by ARTP technology, the mutation rate was 42.8%, the positive mutation rate was 20.6%, and the enzyme activity of G2-1 mutant reached 2.73U/mL, which was 82% higher than that of the original strain.
Fig. 3. Typical colony characteristics after ARTP mutagenesis (G1-G14 was the representative of the typical colony with different morphology from the original strain; G15 was the same as the original strain). (Xia, et al., 2010)
Case Study 3: Screening of monascus mutants with high orange/yellow pigment production using ARTP.
Natural pigments produced by Monascus fermentation have attracted increasing attentions. The optimal irradiation time for the Monascus mutation was 90s, at which the spore lethality rate was 78% and the morphological mutation rate was 14.2%. The highest orange and yellow pigment production was 3,620 and 3,300 U/g by solid-state fermentation of the mutant strain WM951M1 at 30°C for 10d, with an increase of 43% and 136% when compared with that of the original strain.
Fig. 4. The mutant strains with typical morphological changes. (Liu, et al., 2016)
FAQs
Q: Is strain modification applicable to all types of microorganisms or cell lines?
A: Strain modification techniques are applicable to many types of microorganisms and cell lines, but the specific modification strategies and effects may vary depending on strain species and characteristics.
Q: Is the modified strain safe?
A: Strain improvement service providers usually follow strict safety protocols and regulatory requirements to ensure that the modified strains meet standards in terms of safety and environmental impact.
Q: How long does the process of strain improvement take?
A: The duration of the improvement process depends on the complexity of the strain, the specificity of the target characteristics, and the technical means employed, and can range from several days to several weeks.
References:
- Jiang X.; et al. Starting with screening strains to construct synthetic microbial communities (SynComs) for traditional food fermentation. Food Res Int. 2024;190:114557.
- Li H.; et al. Mutagenesis of Methylobacterium extorquens AM1 for increasing pyrroloquinoline quinone production by atmospheric and room temperature plasma. Sheng Wu Gong Cheng Xue Bao. 2016;32(8):1145-1149.
- Xia SQ.; et al. Mutation and Selection of Transglutaminase Producing Strain by Atmospheric Pressure Glow Discharge Plasma. Microbiology China. 2010, 37(11): 1642-1649
- Liu GQ.; et al. Screening of Monascus Mutants With High Orange/Yellow Pigment Production Using Atmospheric and Room Temperature Plasma(ARTP). Journal of Nuclear Agricultural Sciences. 2016, 30(4): 654-661
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