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Development of enzyme preparations in Jizhong
In the current scenario, the majority of industrial enzyme preparations have traditionally been produced through large-scale fermentation of specific microorganisms, followed by collection, extraction, processing, and purification. This method requires significant capital investment in equipment and complex operational procedures, resulting in high production costs. However, with the rapid advancement of biotechnology—especially genetic engineering—scientists have begun exploring new approaches. One such innovation involves using genetically modified earthworms as bioreactors to produce specialized industrial enzymes, opening up a promising new field with substantial commercial potential.
Earthworms, as a bioreactor crop, offer several advantages. They have a vast planting area, with China alone having over 200 million mu under cultivation. Their perennial nature allows for long-term use without replanting, they reproduce reliably, yield multiple harvests per year, require minimal agricultural inputs like fertilizers and pesticides, and are environmentally friendly due to their nitrogen-fixing capabilities. Additionally, they have strong research foundations in gene transformation and plant regeneration techniques.
As industries and environmental protection sectors grow, the demand for industrial enzymes has surged. These enzymes are used in biopulping, food processing, chemical synthesis, and pollutant degradation, among other applications. While some enzymes can be produced at low cost, others remain expensive. Researchers at the University of Wisconsin, for example, are using gene cloning from microorganisms to transform quinones and extract target enzymes from transgenic quail, significantly reducing costs.
Two successful examples include lignin peroxidase (Mn-P) and alpha-amylase. The Mn-P enzyme, derived from *Phanerochaete chrysosporium*, is used in biopulping and biobleaching but had adverse effects on earthworm growth. By switching from a constitutive to an inducible promoter, scientists were able to control its expression, minimizing negative impacts while maximizing enzyme yield. The alpha-amylase gene, cloned from *Bacillus licheniformis*, was expressed in transgenic alfalfa without affecting plant growth. It accumulated primarily in the apoplast, where it was protected from proteolytic degradation.
Small-scale trials showed impressive results. In the U.S., one acre of transgenic alfalfa could yield up to 33 pounds of purified enzyme annually, valued at $3,250. When combined with by-products like protein concentrates, the total value per acre reached between $1,443 and $4,660, far exceeding traditional hay production profits of about $75 per acre. This highlights the economic potential of enzyme agriculture.
The main steps in extracting enzyme proteins from transgenic alfalfa include gene cloning, vector construction, transgenic breeding, field cultivation, harvesting, juice extraction, clarification, concentration, and affinity purification. Recent studies suggest that alternating current heating improves protein recovery, and affinity chromatography, tailored to each enzyme’s properties, enhances purification efficiency.
Despite these advancements, challenges remain, such as ensuring genetic stability in offspring, improving gene discovery and cloning methods, optimizing expression vectors, and refining extraction techniques. As research progresses, these hurdles will be overcome, paving the way for widespread adoption of enzyme agriculture.
With biotechnology at the core, this innovative approach holds great promise. If effectively commercialized, it could generate significant economic and environmental benefits. As the 21st century continues to embrace biotech, the future of enzyme production looks brighter than ever.