Team:NYMU-Taipei/HP/Gold Integrated

We interviewed with professionals to collect their suggestions. We went to the largest microalgae supplier company in Asia, whose CEO mentioned the multi-benefits of algae and encouraged us to tackle different methods to increase algae oil accumulation. This meeting inspired us to include another pathway -- Pigment -- to improve photosynthesis efficiency, thereby producing excess starch that can then be converted to oil. This ended up becoming more effective compared to our initial nitrogen starvation pathway.

Better our algae

We went to Academia Sinica and learned how to maximize algae growth from a plant and microbial professor. We also spoke with Team Tokyo Tech, who inspired us to construct a plasmid containing Melanin (black pigment) sequence for maximizing light intake.

Integrated our testing

We further reviewed the pathway, device, and testing of our project with several professors at National Yang-Ming University. Based on Professor Chen’s advice, we enhance our device (for testing the growth environment) by adding cloud technology. To check the growing situation by measuring OD value, we acquire how to use the Photo-Bioreactor from Dr. Ya-Tang Yang (NTHU team).

Far East Bio Tec

After months of research on increasing the oil production in algae, we started to construct plasmids which would be used in our project. However, some question arose during the process of conducting our project：

Except “denitrification”, is there any other pathway to increase oil production in algae?
How to maximize the potential of algae and make it play a promising role in producing clean energy?
What is the limitation that we haven’t discovered in producing biofuel from algae? Can we make effort to conquer it?

Seek the profession

Besides, we are looking forward to building connections between the society and our lab work. With the eagerness to find out appropriate and feasible answers to these questions , we met with Far East Bio-Tec Company(FEBICO) in their headquarter on July.10 2017. Based on the issues we mentioned in this meeting with FEBICO (silver HP), we dedicated to improve our project in many ways:

(1.) Pigment(Link: Project-Pigment)
This is the most significant change after the meeting with Far East Bio-Tec Company(FEBICO). A brand-new pathway in increasing algae energy production has been created.

(a) High additional value
We learned the importance of additional value in algae. The various products produced by FEBICO inspired us to come up with another pathway ---Pigment---to increase the oil production in algae. With a response to the international market trend, we should develop energy source having multiple functions . As a promising energy source, algae contains crucial nutrients bringing benefits to our human bodies. We are looking forward to presenting the spirit of adding value through the design of pigments in our project.
(b) Part selection
When it came to what pigment we intended to construct, Astaxanthin jumped into our mind since it was mentioned in the meeting with FEBICO. Astaxanthin is a high activity antioxidant and has been suggested being beneficial in human health such as preventing cardiovascular, immune and neurodegenerative diseases.
(c) Another way except “Denitrification”
Compared with “Denitrification” which forces algae to produce more biofuel by making restrictions in the environment, “Pigment” proposes an opposite concept---“ adding conditions ”. We expect that adding pigments in algae can enhance the efficiency of photosynthesis, leading to the increase of oil production. Thanks to FEBICO, we carved out a promising way.

(2.) Device Improvement(Link:Project-Device)
The manufacturing scale of FEBICO stimulates our thoughts to build a device.We are convinced that the introduction of the device into our project can help us gain more precise manipulation, measurement, and response. The design of our device aims to obtain immediate information of algae pond and monitor the growth of microalgae precisely. The business model learned from FEBICO is absolutely a precious material.

(3.) Crack the cell wall
Actually, in the measurement of functional test of NrtA protein, we confront some difficulties at first. Initially, the result showed no significant difference between the treatment group and control group. Inspired by patent of “Cracked Cell wall” proposed by FEBICO, we decided to introduce French Pressure Press into our experiment to extract NrtA protein from the transformants. Further combined with a semi-permeable membrane system, NrtA protein in our experiment was proved valid. Thanks to the advice provided by Mr. Chiueh, the proof of concept was successfully carried out.

(4.) Interest in DNA sequencing
Despite the fact that FEBICO does not have any products made by genomic modification, Mr. C. C. Chiueh (the founder of FEBICO) still showed a great interest in DNA sequencing. DNA sequencing can bring more biological information about algae, leading to profound researches and applications. The largest genomic research center in Taiwan is located in National Yang-Ming University. Mr. Chiueh intended to collaborate with the institute and further build a long-term relationship with NYMU.

Academia Sinica

On July 18th, 2017, to enhance our knowledge on microalgae and seek advice of our project, we visited Academia Sinica, a national academy that supports research in Taiwan. We contacted Professor Chu Hsiu-An, a professor with a Ph.D. in Biochemistry. He works at the Institute of Plant and microbial biology department. He provided us with a supply of Synechocystis species PCC6803 that can survive with its high dependence on glucose. He gave us further information on the optimal conditions for the cultivation of algae. We are really grateful that Dr. Chu was willing to provide us with the algae he uses in his lab. We later cultivated our algae in our own lab.Genomic DNA of PCC6803 was further extracted for the construction of NrtA gene.

Professor Chen

You-Yin Chen is a professor of biomedical engineering a National Yang-Ming University. He is responsible for the integration of semiconductor process, micro-electromechanical technology, and material science, in order to develop a new generation of nerve wafers. Obviously, our device needs his brilliant skills and guidance. Luckily, he is willing to help us set up the initial architecture of the entire project as well as the erection of the network. Also, he provides us rich resources and technology, and guides us through the establishment of the whole program such as pH sensor, motor, circuit design, etc. Furthermore, he teaches us Arduino development version of the application and design. Most importantly, he suggests that we can use the cloud technology to store more data and keep records. Thanks to Professor Chen, we are able to improve our quality of the device.

_{Dr.You-Yin Chen}

National Tsing Hua University

After the 5th Asia-Pacific iGEM Conference, NYMU team had a chance to contact the primary instructor of NTHU team, Dr. Ya-Tang Yang, and his laboratory students. Dr. Yang is the associate professor of Institute of Electronics Engineering, National Tsing Hua University.They provided us with a set of Photo-Bioreactor made by themselves and taught us how to use the Photo-Bioreactor. During August to September, we discussed with Dr. Yang and his lab students, trying to fix some difficulties we encountered.
After the Photo-Bioreactor finally worked smoothly, we started to measure the OD value of microalgae we cultivated, collected huge amounts of data, and drew growth curves for our projects. Meanwhile, we shared collated data to Dr. Yang’s lab for research use. We believe this collaboration would be a great academic exchange and produce some interesting results.

_{Dr. Ya-Tang Yang, NYMU team member with Dr. Yang’s lab}

_{The growth curve of Chlorella vulgaris as well as Synechococcus elongatus PCC 7942}

Mr.Yang

On June 18, 2017, due to the lack of chemicals, NYMU-Taipei team contacted the Teaching Assistant of Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Mr. Nien Chung Yang, who manages the laboratory. Not only did he help us find the required chemicals such as iodine solution, and gradients of breeding medium, BG-11, but also provide us a working operation to prepare the breeding medium. Furthermore, Mr. Yang taught us how to work in experiment safely, and also gave us a lot of experimental advice on lab-working. Thanks to his generous support, we could progress our experiments more smoothly.