Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Plant Biotechnol J. 2019 Jun;17(6):1167-1182. doi: 10.1111/pbi.13122. Epub 2019 Apr 23.
Chemical catalysts are being replaced by biocatalysts in almost all industrial applications due to environmental concerns, thereby increasing their demand. Enzymes used in current industries are produced in microbial systems or plant seeds. We report here five newly launched leaf-enzyme products and their validation with 15 commercial microbial-enzyme products, for detergent or textile industries. Enzymes expressed in chloroplasts are functional at broad pH/temperature ranges as crude-leaf extracts, while most purified commercial enzymes showed significant loss at alkaline pH or higher temperature, required for broad range commercial applications. In contrast to commercial liquid enzymes requiring cold storage/transportation, chloroplast enzymes as a leaf powder can be stored up to 16 months at ambient temperature without loss of enzyme activity. Chloroplast-derived enzymes are stable in crude-leaf extracts without addition of protease inhibitors. Leaf lipase/mannanase crude extracts removed chocolate or mustard oil stains effectively at both low and high temperatures. Moreover, leaf lipase or mannanase crude-extracts removed stain more efficiently at 70 °C than commercial microbial enzymes (<10% activity). Endoglucanase and exoglucanase in crude leaf extracts removed dye efficiently from denim surface and depilled knitted fabric by removal of horizontal fibre strands. Due to an increased demand for enzymes in the food industry, marker-free lettuce plants expressing lipase or cellobiohydrolase were created for the first time and site-specific transgene integration/homoplasmy was confirmed by Southern blots. Thus, leaf-production platform offers a novel low-cost approach by the elimination of fermentation, purification, concentration, formulation and cold-chain storage/transportation. This is the first report of commercially launched protein products made in leaves and validated with current commercial products.
由于环境问题，化学催化剂在几乎所有工业应用中正在被生物催化剂所取代，从而增加了对生物催化剂的需求。目前工业中使用的酶是在微生物系统或植物种子中生产的。我们在这里报告了五种新推出的叶酶产品，并将其与 15 种商业微生物酶产品在洗涤剂或纺织工业中的应用进行了验证。叶绿体中表达的酶作为粗提叶提取物，在广泛的 pH/温度范围内具有功能，而大多数纯化的商业酶在碱性 pH 或更高温度下会显著失活，这是广泛应用所必需的。与需要冷藏/运输的商业液体酶不同，作为叶粉的叶绿体酶可以在环境温度下储存长达 16 个月而不会失去酶活性。叶绿体衍生的酶在粗提叶提取物中是稳定的，无需添加蛋白酶抑制剂。叶脂肪酶/甘露聚糖酶粗提取物可有效去除巧克力或芥末油污，无论在低温还是高温下。此外，叶脂肪酶或甘露聚糖酶粗提取物在 70°C 下比商业微生物酶（<10%活性）更有效地去除污渍。粗叶提取物中的内切葡聚糖酶和外切葡聚糖酶可有效去除牛仔布表面和起绒针织物的染料，通过去除水平纤维束。由于食品工业对酶的需求增加，首次创建了表达脂肪酶或纤维二糖水解酶的无标记生菜植物，并通过 Southern 印迹证实了基因定点整合/同型性。因此，叶片生产平台通过消除发酵、纯化、浓缩、配方和冷链储存/运输，提供了一种新颖的低成本方法。这是首次报道在叶片中生产并与当前商业产品进行验证的商业推出的蛋白质产品。
