GOALI: E. coli Metabolic Engineering to Produce the Marine Anticancer Agent Lomaiviticin A

0932612 Pfeifer智力价值这个项目源于塔夫茨大学和惠氏研究之间的互补伙伴关系。Wyeth Natural Products Discovery Group被公认为工业天然产物药物发现和开发的领导者，最近对一种名为Lomaiviticin A的强效抗癌化合物的基因簇进行了测序。该基因序列分离自天然负责洛麦维他菌素A生产的海洋细菌太平洋盐孢菌。不幸的是，S. pacifica是耗时的（7-14天），不一致，低产量（0.5毫克/升）。在这个项目中，E.大肠杆菌将被用作洛麦替星A生物合成的异源宿主，其逻辑是E.大肠杆菌将克服目前的S.太平洋生产的局限性。在这里，塔夫茨大学的主要研究者（PI）小组将努力将洛麦维菌素A基因簇转移到大肠杆菌中，以进行后续的异源生产。Tufts团队专门研究遗传转移策略，以通过方便的异源宿主促进复杂的天然产物生产。总之，工业-学术合作非常适合通过理想的生物宿主生产这种重要的海洋抗癌剂。更广泛的影响塔夫茨和惠氏之间的协同和互补的学术工业伙伴关系将促进独特的更广泛的影响机会。通过塔夫茨大学，该项目将利用研究作为教育和外联机制。研究本身跨越了许多学科，包括：分子生物学，微生物学，化学，遗传学，化学和生物工程。两个教育水平（研究生和本科）的学生将有机会参与并学习这种多学科方法。此外，个人研究责任的目的是重新加强学生的学术水平。研究生将监督和管理目标1和2的实验工作，同时指导本科生助理。因此，研究生将发展成为一名独立的科学家，并将获得指导经验，因为他/她训练年轻的学生。本科生将学习过程培养和分析技能，符合他们的化学和生物工程课程。作为推广计划的一部分，本科生将从塔夫茨NSF赞助的计算机科学，工程和数学学者计划中选出，该计划鼓励代表性不足的群体追求科学和工程，PI将担任学术顾问。 最后，学生将能够比较学术和工业研究机会，并通过惠氏主办的暑期实习获得第一手资料。

0932612PfeiferIntellectual merit This project originates from a complementary partnership between Tufts University and Wyeth Research. The Wyeth Natural Products Discovery Group is recognized as a leader in industrial natural products-based drug discovery and development and has recently sequenced the gene cluster responsible for a potent anticancer compound called lomaiviticin A. The gene sequence was isolated from the marine bacterium Salinispora pacifica naturally responsible for lomaiviticin A production. Unfortunately, lomaiviticin A production from S. pacifica is time-consuming (7-14 days), inconsistent, and low-yielding (0.5 mg/L). In this project, E. coli will be used as a heterologous host for lomaiviticin A biosynthesis with the logic that E. coli will overcome the current S. pacifica production limitations. Here, the Principal Investigator's (PI's) group at Tufts University will lead efforts to transfer the lomaiviticin A gene cluster to E.coli for subsequent heterologous production. The Tufts team specializes in genetic transfer strategies to facilitate complex natural product production through convenient heterologous hosts. Together, the industrial-academic collaboration is well-suited for the production of this important marine anticancer agent through perhaps the ideal biological host. Broader impactsThe synergistic and complementary academic-industrial partnership between Tufts and Wyeth will promote unique broader impact opportunities. Through Tufts, the project will use research as an educational and outreach mechanism. The research itself crosses numerous disciplines that include: molecular biology, microbiology, chemistry, genetics, and chemical and biological engineering. Students at two educational levels (graduate and undergraduate) will have the opportunity to participate in and learn from this multi-disciplinary approach. In addition, individual research responsibilities have been designed to re-enforce the academic level of the student. The graduate student will oversee and manage the experimental efforts in Aims 1 and 2 while simultaneously guiding undergraduate assistants. Hence, the graduate student will develop as an independent scientist and will also gain mentoring experience as he/she trains younger students. The undergraduate student will learn process culturing and analysis skills that coincide with their chemical and biological engineering curriculum. As part of the outreach initiatives, the undergraduate will be selected from the Tufts NSF-sponsored Computer Science, Engineering, and Mathematics Scholars program that encourages underrepresented groups to pursue science and engineering and for whom the PI serves as an academic advisor. Finally, the students will be able to compare academic and industrial research opportunities and gain first-hand access to both climates through summer internships hosted by Wyeth.