Bioengineering a non-pathogenic bacteria to produce medically-relevant biopolymers

项目摘要/摘要 藻酸盐是一种生物聚合物，在食品、个人护理产品和生物医药中有着广泛的应用。在……里面 除了作为片剂赋形剂和抗酸治疗的作用外，海藻酸盐还作为一种非 粘附性渗出物吸收凝胶，用于高级伤口护理敷料。从历史上看，这是一种藻类 已经从棕色海藻中获得了应用，但这种来源的问题包括减少 产量、需求大于供应，以及不能改变的固定聚合物组成 生产独特的定制藻酸盐，以加强伤口闭合和防止感染。某些种类的 细菌(固氮菌、假单胞菌)产生海藻酸盐，但由于海藻酸盐的减少而阻碍了商业化 或在培养中不稳定地合成这种聚合物。突破性和专利技术是由 Progensis的联合创始人，于宏伟博士，涉及一种基因信号，以激活高产和稳定的生产 一种工程菌株铜绿假单胞菌。这个SBIR项目的产品将是不同单体的海藻酸盐 不是由海藻产生的组合物和修饰(乙酰化)。这些海藻酸盐将产生 钠或钙凝胶具有不同的抗张强度、孔径、液体保留率和药物释放速率，可以 可定制以提高其应用于藻酸盐创面敷料的有效性。在第一阶段 项目中，一株铜绿假单胞菌pgn5被改造成缺乏编码相关产物的基因。 对FDA来说是必不可少的致病因素。PGN5在小鼠中无致病性，类似于FDA批准的大肠杆菌 K12。在这个第二阶段的提案中，Progensis公司将进一步对PGN5菌株进行生物工程，以生产 会形成不同抗张强度、孔隙率和吸液能力的凝胶，使藻酸盐伤口 敷料可以根据每个病人的需要进行个性化(目标1)。目标2将确定物理化学 与海藻藻酸盐敷料相比，将伤口闭合时间缩短20%所需的特性。为了确保 充足的供应和利润率Progensis将对中央碳代谢进行生物工程，以增加 目前藻酸盐的产量提高了50%，并扩大到45L生物反应器(目标3)。实现这些目标将使 产生独特的藻酸盐，以促进慢性伤口的愈合，据报道 2014年全球市场规模达99亿美元。含有藻酸盐的敷料约占市场的5%-7%， ~6.9亿美元。它们也是创伤护理的组成部分，预计年复合增长率最高 到2021年，税率(16%)。临床前数据将用于制定包含里程碑的研发协议 向销售伤口护理产品的公司付款。此外，还将寻求与 生产海藻藻酸盐的公司从特许权使用费中提供了额外的现金流。前生生物也将 向研发实验室推销其藻酸盐套件，用于干细胞等各种研究应用 分化、3-D癌细胞生长和组织工程学。这将在研发期间提供收入流 并通过谈判达成许可协议。

Project Summary/Abstract Alginate is a biopolymer with a wide array of applications in, food, personal care products and biomedicine. In addition to its role as a tablet excipient and antacid treatment, alginates play an important role as a non- adherent, exudate absorbing gel, for advanced wound care dressings. Historically, alginates for this application have been obtained from brown seaweed, but the problems with this source include decreasing yield, demand that is larger than the supply, and a fixed polymer composition that cannot be changed to produce unique alginates customized to enhance wound closure and prevent infections. Certain species of bacteria (Azotobacter, Pseudomonas) produce alginate, but commercialization has been blocked by decreased or unstable synthesis of this polymer in culture. Breakthrough and patented technology was developed by Progenesis co-founder, Dr. Hongwei Yu that involves a genetic signal to activate high and stable production in an engineered strain of P. aeruginosa. The product of this SBIR project will be alginates of different monomer compositions and modification (acetylation) that are not produced by seaweed. These alginates will produce sodium or calcium gels with varying tensile strength, pore size, fluid retention and drug release rates that can be customized for improving the effectiveness of their application for alginate wound dressing. In the Phase I project, a strain of P. aeruginosa, PGN5 was engineered that lacked the genes encoding products of concern to the FDA and essential to pathogenesis. PGN5 is non-pathogenic in mice similar to the FDA-approved E coli K12. In this phase II proposal Progenesis will further bio-engineer the PGN5 strain to produce alginates that will form gels of different tensile strength, porosity and fluid adsorption capability so that alginate wound dressings can be individualized for each patient’s needs (aim1). Aim 2 will determine the physical-chemical properties necessary to reduce wound closure time by 20% relative to seaweed alginate dressings. To ensure adequate supply and profit margin Progenesis will bio-engineer central carbon metabolism to increase the current yield of alginates by 50% and scale up to a 45L bioreactor (aim 3). Achieving these aims will allow Progenesis to produce unique alginates tailored to enhance the healing of chronic wounds with a reported 2014 global market of $9.9 billion USD. Alginate containing dressings comprise about 5-7% of the market, ~$690 million USD. They are also the component of wound care that has the highest projected CAGR growth rate (16%) through 2021. The preclinical data will be used to develop R&D agreements containing milestone payments with companies selling wound care products. Also licensing agreements will be sought with companies producing seaweed alginate providing additional cash flow from royalties. Progenesis will also market its suite of alginates to R&D laboratories for use in various research applications such as stem cell differentiation, 3-D cancer cell growth, and tissue engineering. This will provide a revenue stream while R&D and licensing agreements are negotiated.