Biodiversity in Vietnam leads the industry forward

It all started ten years ago when Vu Nguyen Thanh, a researcher at the Food Industries Research Institute, FIRI, in VIetnam emailed Lisbeth Olsson, Professor and Head of the Division of Industrial Biotechnology, to discuss a research collaboration. The Swedish Embassy in Hanoi announced a call for research grants to promote collaboration between Swedish and Vietnamese researchers wanting to solve local environmental problems.

Efficient enzymes the key to larger-scale production

At that time, Vu Nguyen Thanh's focus was to produce bioethanol from residues from the agricultural industry, in the form of lignocellulose.

“We had come to the conclusion that the key to a larger-scale production was enzymes that effectively help break down lignocellulose, and we knew about Lisbeth Olsson’s work in that field,” says Vu Nguyen Thanh.

Lisbeth Olsson is professor of industrial biotechnology and her research aims to design enzymes and microorganisms into processes where plant cell wall material is converted into fuel, chemicals and biomaterials. She is also the Vice Director of the Wallenberg Wood Science Center (WWSC), a research center focused on research to create future high-tech materials from wood.

Too expensive to buy enzymes

“At first, I received some emails from Thanh, that I did not respond to right away. When he called me and explained more, I could see the project’s potential. We applied for, and received, the current grant. Finding native solutions, microorganisms and enzymes is crucial in countries like Vietnam,” says Lisbeth Olsson and continues:

“Enzymes are expensive, and they cannot afford to buy the material they need for these processes. In addition, there can be many other benefits of producing the enzymes in Vietnam, as they can be adapted to their local needs.”

Enzymes can contribute to reducing​ environmental effects

These enzymes could be of great benefit in other areas besides biofuel production. There is also a great need in the animal feed industry to increase the nutritional value. In addition, environmental problems in agriculture, among other examples, need to be addressed.

“Vietnam is largely an agricultural country, and we are, among other things, one of the world's largest rice exporters. After the harvest, the residual products, i.e. the rice straw, are often burned to make space for new crops and to recycle minerals. It causes pollution and wastes the resources that could otherwise be converted into valuable products, such as animal feed, but also bioethanol and chemicals. For such products to be created on a large scale and cost-efficient, enzymes are needed,” says Vu Nguyen Thanh.

Unexplored biodiversity ​in the rainforest

The Vietnamese rainforests show a great biodiversity and there are many microorganisms that effectively decompose dead plant material. In the diversity of the rainforest there are great opportunities for enzyme discoveries, but this biodiversity is still mostly unexplored.

FIRI's ambition was to collect and analyse domestic microorganisms with the potential to be used industrially in the future, but greater knowledge in the area was required. Therefore, Vietnamese research students visited Chalmers to learn more about molecular biological methods, fermentation and analysis, but also how to design experiments and interpret results.

Collaboration good for both parties

Johan Larsbrink, Associate Professor, and Silvia Hüttner, formerly postdoc, at the Division of Industrial Biotechnology, have participated in the training of the Vietnamese researchers. They have also visited Vietnam several times with Lisbeth Olsson to hold workshops, network with researchers and visit both smaller and larger local industries, and of course participate in the collection of microorganisms. The Chalmers researchers believe that the collaboration with FIRI have benefited both parties.

“I believe it was rewarding for the students who came to Chalmers to work in a whole new environment. And for us it was very good to get in touch with the researchers in Vietnam to gain access to their biodiversity and their knowledge about it. Both parties in the collaboration gained new experiences and access to knowledge or biomaterials. In our case, we discovered some interesting organisms that we are still analysing and that can lead to more projects and results in the future,” says Silvia Hüttner.

​"FIRI wants immediate impact"

Johan Larsbrink agrees that the collaboration has been a good experience, both in the collection of the microorganisms, but also in the laboratory.

“It is interesting to see how they use the resources they have, which are limited. Although the Chalmers group wants to see applications and results of our research in the community, we are also conducting ‘curiosity-driven research’, while FIRI has a greater interest in the research having immediate impact and being used in society directly,” he says.

Industry and research benefit from the discoveries

Vu Nguyen Thanh says that he personally has been able to better picture what can be done to make use the microbial resources that exist in the country. The focus has always been on local industry and local companies being able to benefit from discoveries and innovations that have sprung from the collaboration. And they have been able to do just that. During the ten years of joint research, the collaboration has been awarded more grants and produced results.

An example is the enzyme xylanase used in animal feed. It is added to feed pellets for chickens and is activated when it reaches the animal's digestive tract. The enzyme helps the chicken to digest the dietary fiber in the feed and thus raises the nutritional value for the animal, compared to feed where the enzyme has not been added.

Several good examples​

Trichoderma harzarium is another example. It is a fungus that can be used in so-called vermicomposting, composting using worms, for more efficient degradation of rice straw. When the compost is used as fertilizer, the fungus also protects plant roots from various diseases.

“We are currently also investigating the possibility of using fungi to convert organic residues from wastewater after ethanol distillation into fungal biomass that can be used for animal feed,” says Vu Nguyen Thanh.

“Impressed by the swift process from innovation to production"​

Lisbeth Olsson is happy and proud that Chalmers researchers have also participated in creating forums for local companies and researchers, for example through workshops, where new, local networks have been created.

“I am impressed by the swift process from innovation to production that we have witnessed. I also think it has been very interesting to see how Vietnam has developed during the years we have had contact with FIRI and how fast the development has been for the country. Personally, I also think it has been very satisfying to see that our joint efforts clearly have contributed to their development,” she says.

New research grant −​ the collaboration continues

In December 2020, Lisbeth Olsson and Johan Larsbrink were awarded a research grant from the Swedish Research Council for the three-year project "Exploring fungal extremophiles in South East Asian biodiversity as the key for sustainable agriculture and industry", thus the successful collaboration will continue.

“In collaboration with our colleagues, we will explore the biodiversity in South East Asia for useful enzyme,” Johan Larsbrink explains.

“We will use both acacia bark and cassava residues from the bioethanol industry and enzymes that rapidly degrades and valorize these materials,” says Lisbeth Olsson.

The long-term goal with the project is to create valuable products from the biomass resources, which today are discarded in huge quantities and as a consequence cause pollution issues. New transformation methods could create great value and benefit local industries in the region, and in the long run, could also lift even more people out of poverty.

New or improved products, services and processes that led to the Chalmers / FIRI collaboration

• Development of a recombinant strain of the yeast Pichia pastoris, which produces a thermostable variant of the enzyme xylanase, which is also tolerant to acid and pepsin. Due to its high thermostability, the enzyme can be preserved by spray drying instead of freeze drying. This improvement makes enzyme production more cost-effective.
• FIRI supplies starting cultures of Aspergillus and Rhizopus fungi to local alcohol fermentation companies, and lignocellulolytic strains to universities (Hanoi University of Technology, Vietnam National University of Agriculture and Hanoi National University of Science) for research and educational purposes.
• FIRI helps bioethanol producers in Vietnam to use lignocellulolytic enzymes to reduce the viscosity of cassava fermentation. Getting more fluid-flowing processes makes it easier to handle the large volumes involved and can also increase the fermentation rate.
• FIRI has become one of the leading institutions in Vietnam in enzyme technology and has produced several enzymes of industrial interest (for example, phytase and sucrose isomerase). FIRI works with the sugar manufacturer LASUCO in the demonstration phase for isomaltose production where a recombinant sucrose isomerase is used. FIRI also offers analytical services for analysing enzyme activities and identifying microorganisms with DNA sequencing for local industry.

The collaboration between FIRI and Chalmers has resulted in the following publications

• Specific xylan activity revealed for AA9 lytic polysaccharide monooxygenases of the thermophilic fungus Malbranchea cinnamomea by functional characterization
• Surveying of acid-tolerant thermophilic lignocellulolytic fungi in Vietnam reveals surprisingly high genetic diversity​​
• Genome sequence of Rhizomucor pusillus FCH 5.7, a thermophilic zygomycete involved in plant biomass degradation harbouring putative GH9 endoglucanases
• Combined genome and transcriptome sequencing to investigate the plant cell wall degrading enzyme system in the thermophilic fungus Malbranchea cinnamomea​