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Construction and Evaluation of High-Efficiency Tannase-Producing Strains
Yuan Gao1, Chenguang Hu1, Wurilege Wei1
1Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Hohhot 010031, China.
Microorganisms
|June 26, 2026
Summary
This study engineered Bacillus subtilis strains for enhanced tannase production, addressing low enzyme efficiency and poor utilization of tannin-rich feed. The WB600(pHT43) strain showed superior stability and enzyme output, advancing industrial enzyme production.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Microbial Fermentation
Background:
- Low production efficiency of tannase limits its industrial application.
- Inefficient utilization of high-tannin feed resources necessitates improved enzyme solutions.
- Tannase enzyme plays a crucial role in degrading tannins, improving feed quality.
Purpose of the Study:
- To develop efficient tannase-producing strains of Bacillus subtilis through heterologous expression.
- To evaluate the growth characteristics, expression stability, and enzymatic properties of recombinant strains.
- To identify the optimal strain for industrial tannase production and feed utilization.
Main Methods:
- Gene cloning and heterologous expression of tannase (TanLpl) from Lactiplantibacillus plantarum in Bacillus subtilis 168 and WB600.
- Construction of recombinant strains using plasmids TanLpl-p43NMK and TanLpl-pHT43.
- Assessment of growth kinetics, plasmid stability through subculturing, and enzyme activity under varying conditions (temperature, pH).
Main Results:
- Three recombinant tannase-producing strains were successfully constructed: BS168(p43NMK), BS168(pHT43), and WB600(pHT43).
- All strains exhibited similar growth rates to parental strains, reaching stationary phase at 18 h.
- WB600(pHT43) demonstrated superior expression stability and significantly higher tannase activity (461.12 U/mL) at optimal conditions (30 °C, pH 5.0) compared to BS168(pHT43) (397.36 U/mL) and BS168(p43NMK) (68.81 U/mL).
- Plasmid loss rate was significantly lower in BS168(pHT43) and WB600(pHT43) compared to BS168(p43NMK) after 10 generations.
Conclusions:
- Heterologous expression of tannase in Bacillus subtilis is feasible and can significantly enhance enzyme production.
- The recombinant strain WB600(pHT43) is a promising candidate for industrial-scale tannase production due to its high activity and stability.
- This study contributes to the technical system for functional enzyme heterologous expression and promotes bio-manufacturing technology for feed utilization.
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