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

该项目源于塔夫茨大学和惠氏研究公司的互补合作伙伴关系。惠氏天然产物发现小组是公认的工业天然产物药物发现和开发的领导者，最近对一种有效的抗癌化合物lomaiviticin a的基因簇进行了测序，该基因序列是从海洋细菌Salinispora pacifica中分离出来的，该细菌自然负责lomaiviticin a的生产。不幸的是，从太平洋杉中生产洛马维菌素A耗时（7-14天），不稳定，产量低（0.5 mg/L）。在本项目中，大肠杆菌将作为lomaiviticin a生物合成的异源宿主，其逻辑是大肠杆菌将克服目前太平洋葡萄球菌的生产限制。在这里，塔夫茨大学的首席研究员（PI）小组将领导将lomaiviticin A基因簇转移到大肠杆菌中进行后续的异源生产。塔夫茨团队专门研究遗传转移策略，通过方便的异源宿主促进复杂的天然产物生产。总之，这种工业-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-产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，该项目将把研究作为一种教育和推广机制。这项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。两个教育层次（研究生和本科生）的学生将有机会参与并从这种多学科方法中学习。此外，个人研究责任的设计是为了加强学生的学术水平。研究生将监督和管理目标1和目标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，并在他/她培训年轻学生的过程中获得指导经验。本科学生将学习过程培养和分析技能，与他们的化学和生物工程课程相吻合。作为拓展计划的一部分，该本科生将从塔夫茨国家科学基金会（Tufts nsf）赞助的计算机科学、工程和数学学者项目中选出，该项目鼓励未被充分代表的群体攻读科学和工程，PI将为他们担任学术顾问。最后，学生们将能够比较学术和工业研究机会，并通过惠氏主办的夏季实习获得两种气候的第一手资料。

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.