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11 July 2022 | Story Andre Damons | Photo Supplied
Prof Martie Smith and Prof Drik Opperman
Prof Martie Smit and Prof Dirk Opperman in the Department of Microbiology and Biochemistry filed a patent entitled “Process for the chemical modification of alkanes, fatty acids and fatty alcohols”.

Flavours and fragrances have a wide application in the food, feed, cosmetic, chemical and pharmaceutical sectors. Many flavour compounds are still produced via chemical synthesis or via extraction from plant or animal sources. However, there is increasing interest in their bio-production or the use of flavour compounds of (micro) biological origin. 

One reason for this shift is that chemical synthesis often uses environmentally unfriendly processes. Chemical synthesis usually also produces racemic mixtures with the second enantiomer, mirror image of the looked-for compound, often having undesirable organoleptic properties. Furthermore, the consumer has developed a “chemophobia”-attitude towards synthetic chemical compounds, especially when related to food and home-care products.  This applies even to nature-identical compounds – products that occur in nature but are produced via a non-natural chemical process. Products produced with the use of enzymes or microbes from “natural” substrates can be labelled “natural”. The flavour and fragrance industry thus pay higher prices for such products labelled as “natural”.  

The invention

A University of the Free State (UFS) team, led by Prof Martie Smit and Prof Dirk Opperman in the Department of Microbiology and Biochemistry are conducting exciting research in this area. They filed a patent entitled “Process for the chemical modification of alkanes, fatty acids and fatty alcohols”.  

The invention relates to a process for the enzymatic in-chain hydroxylation of C12 to C16 fatty acids, alcohols, and alkanes. Hydroxylation of C12 fatty acid and alcohol provides routes for the synthesis of “natural” δ-dodecalactone. The advantage of these routes is that they do not rely on massoia lactones. Massoia lactones are derived from the bark of Massoia trees which grow in Indonesia. Harvesting of the bark kills the trees.  

The cytochrome P450 enzymes (P450s) claimed in this patent are to the inventors’ knowledge the most regioselective enzymes described thus far that can be used for the synthesis of δ-dodecalactone from lauric acid or 1-dodecanol. The approach that the technology takes is to claim cytochrome P450 enzymes that share 70 % amino acid identity to a set of selected P450s for the regioselective hydroxylation of lauric acid and 1-dodecanol to synthesise δ-dodecalactone.

Still in early stage

The current state of development is early stage with the technology only demonstrated in the laboratory on a small scale (100-200 ml). Before the technology can be commercialised the team would need to further improve the regioselectivity and stability of the P450s and proof that the reactions can be scaled up in bioreactors. The technology will probably be delivered as an enzyme (amino acid sequence) with the desired properties. 

There are other research groups working on a synthetic biology approach for the de novo synthesis of δ-dodecalactone from glucose by genetically engineered microbes. It is still unclear how such a process will compare in terms of product yields, economics and environmental impact with the processes proposed by the UFS patent.

If the team had to partner with a commercial company, their first choice would be to work with an established flavour and fragrance company. Another possibility would be the small French flavour and fragrance company that Dr Alizé Pennec, the post-doc and co-inventor who initially discovered the unique P450 activity, is working for.

Please view the videos for more information on patents.

The Vice-Rector: Research and Internationalisation has released two new calls for applications for funding. Academic staff and researchers are encouraged to submit applications for these funds. At this stage we are not accepting projects from Research Fellows. 

The two funds are: 

1.  The Industrial Engagement Fund 
2.  The Intellectual Property Commercialisation Fund

Each fund has its own guidelines and application process. The guidelines are attached. The applications must be filled in on RIMS.

The RIMS application forms can be found through this link

For more information please click the documents below:



News Archive

Research by experts published in Nature
2011-06-02

 
The members of the research group are, from the left, front: Christelle van Rooyen, Mariana Erasmus, Prof. Esta van Heerden; back: Armand Bester and Prof. Derek Litthauer.
Photo: Gerhard Louw

A  research article on the work by a team of experts at our university, under the leadership of Prof. Esta van Heerden, and counterparts in Belgium and the USA has been published in the distinguished academic journal Nature today (Thursday, 2 June 2011).

The article – Nematoda from the terrestrial deep subsurface of South Africa – sheds more light on life in the form of a small worm living under extreme conditions in deep hot mines. It was discovered 1,3 km under the surface of the earth in the Beatrix Goldmine close to Welkom and is the first multi-cellular organism that was found so far beneath the surface of the earth. The worm (nematode) was found in between a rock face that is between 3 000 and 12 000 years old.

The research can shed some new light on the possibility of life on other planets, previously considered impossible under extreme conditions. It also expands the possibilities into new areas where new organisms may be found.

These small invertebrates live in terrestrial soil subjected to stress almost for 24 hours They live through sunshine, rain, scorching temperatures and freezing conditions. Through time they developed a means to cope with harsh conditions. Terrestrial nematodes (roundworms, not to be confused or related to earthworms) are among those very tough small invertebrates that deal with those conditions everywhere. After insects they are the most dominant multi-cellular (metazoan) species on the planet having a general size of 0,5 to 1 mm and are among the oldest metazoans on the planet, Nature says in a statement on the article.

They inhabit nearly every imaginable habitat form the deep seas to the acid in pitcher . Some nematodes simply eat bacteria and these are the ones we study here. Terrestrial nematodes have developed a survival stage that can take them through hard times (absence of food, extreme temperatures, too little oxygen, crowding, and more).

At the head of the research was Prof. Gaetan Borgonie of the Ghent University in Belgium and a world leader in the discipline of nematode research. He was brought into contact with the South African research leader, Prof. Esta van Heerden, who set up a cooperation agreement with the University of Ghent and Prof. Borgonie. Prof. Van Heerden manages the Extreme Biochemistry group at the UFS and the research was funded by several research grants.

The search for worms began in earnest in 2007, but it was soon clear that the sampling strategy was insufficient. A massive sampling campaign in 2008-2009 in several mines led to the discovery of several nematodes and the new nematode species Halicephalobus mephisto. It is named after the legend of Faust where the devil, also known as the lord of the underworld is called Mephistopheles.

Nature says special filters had to be designed and installed on various boreholes. Unfortunately, there is no easy way of finding a magic formula and designs had to be adapted by trial and error; improving existing designs all the time. The work of the UFS Mechanical Workshop, which manufactured, adapted and helped design it, was crucial in this respect. Filters were left on the holes for varying periods, sometimes for a few hours and sometimes for months. Prof. Derek Litthauer from the UFS played a big role in sampling, filter designs and coming up with ideas for names for the new nematode with Prof. Borgonie.

Research showed that the nematodes can live in the deep for up to 12 000 years. Three students – Armand Bester, Mariana Erasmus and Christelle van Rooyen from the UFS – did the work on this.

The importance of multi-cellular animals living in the ultra-deep subsurface is twofold: The nematodes graze on the existing bacterial population and influence their turnover. Secondly, if more complex multi-cellular organisms can survive in the deep subsurface on earth, this may be good news when looking for life on other planets where the surface is considered too inhospitable (e.g. Mars). Complex life forms can be found in ecosystems previously thought to be uninhabitable. Nature says this expands the possibilities into new areas where new organisms may be discovered.

Future research will focus on selective boreholes to look for more metazoans, so that a better idea of the complexity of the ecosystems there can be obtained. It will also look for metazoans in the deep subsurface on other continents to determine similarities and differences.

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