Botryococcus braunii

Botryococcus braunii (Bb) is a green, pyramid-shaped planktonic microalga that is of potential importance in the field of biotechnology . Colonies held together by a lipid biofilm matrix can be found in tropical gold or oligotrophic lakes and estuaries, and will bloom when in the presence of elevated levels of inorganic dissolved phosphorus. The species is notable for its ability to produce high amounts of hydrocarbons , especially in the form of triterpenes , which are typically around 30-40% of their dry weight. [1]Compared to other green algae species it has a relatively thick cell wall that is accumulated from previous cellular divisions; making of cytoplasmic components rather difficult. Fortunately, much of the useful hydrocarbon oil is outside of the cell. [2]

Optimal growth environment

Botryococcus braunii has been shown to grow at a temperature of 23 ° C, a light intensity of 60 W / m², with a light period of 12 hours per day, and a salinity of 0.15 molar NaCl. [3]However, this was the results of testing with one strain, and others certainly vary to some degree. In the laboratory, B. braunii is commonly grown in cultures of Chu 13 medium.

Toxic blooms and competition

Blooms of Botryococcus braunii have been shown to be toxic to other microorganisms and fishes. The cause of the blooms and their subsequent damage to the populations of other organisms has been studied. The exudate of Bb in the form of free fatty acids has been identified as the cause. A higher alkalinity changes these free fatty acids into one form, which causes Bb to become more dominant. Higher alkalinity often occurs when the body is covered with water. While the dominance of Bb can be seen as damaging to the environmental diversity of a body of water, the knowledge of how can be gained and maintained. quote needed ]

Biofuel applications of Botryococcus oils

Main article: Biofuel applications of botryococcene

The practice of farming is known as algaculture . Botryococcus braunii has great potential because of the hydrocarbons it produces, which can be chemically converted into fuels. Up to 86% of the dry weight of Botryococcus braunii can be long-chain hydrocarbons. [4] The vast majority of these hydrocarbons are botryocuccous oils: botryococcenes, alkadienes and alkatrienes. Transesterification can not be used to make biodiesel from Botryococcusoils. This is because these oils are not essential oils in the common sense, in which they are fatty acid triglycerides . Whereas Botryococcus oils are oils of vegetable origin, they are inedible and chemically very different, being triterpenes , and lacking oxygen atom needed for transesterification. Botryococcus oils can be used as feedstock for hydrocracking in an oil refinery to produce octane ( gasoline , aka petrol), kerosene , and diesel . [5] (see vegetable oil refining ). Botryococcenes are preferred over alkadienes and alkatrienes for hydrocracking as botryococcenes will likely be converted into a fuel with a higher octane rating .


Three major breeds of Botryococcus braunii are known, and they are distinguished by the structure of their oils. Botryococcenes are unbranched isoprenoid triterpenes having the formula C n H n -10 . The A race Produces alkadienes and alkatrienes (derivatives of fatty acids ) où n is an odd number 23 through 31. The B race Produces botryococcenes où n is in the ranks 30 through 37 of biofuels choice for hydrocracking to gasoline-like hydrocarbons. The “L” strain makes an appearance of other strains of Botryococcus braunii. Within this major classification, various strains of Botryococcus will differ in the precise structure and concentrations of the hydrocarbon oils. [6]

According to page 30 on the Aquatic Species Program (ASP) report, [7] the A-strain of Botryococcus would have a feedstock for lipid-based fuel production due to its slow growth (one doubling every 72 hours). However, subsequent research by Qin showed that the doubling time could be reduced to 48 hours in its optimal growth environment. [3] In view of findings by Frenz, [6] the doubling times may be important to the method of hydrocarbon harvest. The ASP also found A-strain Botryococcus oil to be less than ideal, having most of its lipids as C 29 to C 34 aliphatic hydrocarbons, and less abundance of C 18fatty acids. This evaluation of Bb oils was done in relation to their suitability for transesterification (ie creating biodiesel ), which was the focus of the ASP at the time Bb was evaluated. The ASP did not study Bb oils for their suitability in hydrocracking , as some subsequent studies were done on the “B” race.

Hydrocarbon Oil Constituents of Botryococcus braunii [5]
Compound  % mass
Isobotryococcene 4%
Botryococcene 9%
34 H 58 11%
36 H 62 34%
36 H 62 4%
37 H 64 20%
Other hydrocarbons 18%

The two listed C 36 H 62 entries are not typos; they are for two different isomers

Extraction of Oils

Compared to other green algae species, Botryococcus braunii has a relatively thick cell wall that is accumulated from previous cellular divisions; making of cytoplasmic components rather difficult. Fortunately, much of the useful hydrocarbon oil is outside of the cell. [8] acting as a biofilm to aggregate individual cells into colonies. The best method of separating the oils from the cells. For some time, it has been known that hexanecan perform this function. However, an electrical method can be cleaner and better overall. Electric fields have been applied in short pulses to hydrocarbon extract from other species of microalgae by weakening the cell walls. These pulses have been microseconds to milliseconds in length. In April 2017 it was reported [9]Researchers at Kumamoto University in Japan have used shorter, nanosecond long pulses to target the extracellular matrix of Botryococcus braunii. They found the method to be less costly and less damaging to the cells than other methods. The Kunamoto scientists found that when the pulses are applied, the optimal field strength was 50 kilovolts per centimeter and the optimum energy applied to 55.6 Joules per milliliter of Botryococcus braunii matrix. Unfortunately, polysaccharides are also extracted from the matrix and must be separated from the oils.


Due to the burgeoning interest in alternatives to fossil fuels , there has been renewed research interest in Botryococcus braunii . The DOE Joint Genome Institute is sequencing the DNA of Bb in 2009-2010. [10] B. braunii is also the subject of research into the production of butylated hydroxytoluene (BHT), [11] an antioxidant, food additive, and industrial chemical.

Potentially useful strains

This heading is a collection of strains of note because of their potential utility. Some of these strains are patented as a result of active DNA modification; furthermore, others are from traditional selection processes.

In 1988, UCBerkeley was granted US Plant Patent 6169 for Botryococcus braunii variety Showa , developed by UC Berkeley scientist Arthur Nonomura, in the Melvin Calvin Laboratory as part of the Nobel laureate’s groundbreaking interdisciplinary program for the development of renewable transportation fuels. The proprietary variety was notable, says the patent application, because of its highly reproducible botryococcal hydrocarbon content including 20% ​​of the dry weight of “Showa.” It is clear that it has been listed as a source of hydrocarbons of its time. The patent expired in April 2008.

In May 2006, Nonomura filed an international patent application disclosing novel growth and harvesting processes for Chlorophyta. [12] A separate patent for plants est is filed Botryococcus braunii variety Ninsei That exhibits the feature of extracolonial secretion of it botryococcenoids That can be processed in refineries Existing gasoline to transportation fuels.

In August 2011, Enomoto was announced by IHI NeoG Algae LLC. [13] It has ” … the highest yield for this fuel production in the world that has been discovered in the world “, with a normal growth rate of more than normal Bb strains. It is more likely to be very robust, [14] presumably meaning it could be grown in an open environment (in ponds, instead of photobioreactors).

See also

  • Torbanite , a form of Botryococcus braunii deposits


  1. Jump up^ Metzger, P .; Largeau, C. (2005). “ Botryococcus braunii : a rich source for hydrocarbons and related ether lipids”. Applied Microbiology and Biotechnology . 66 (25): 486-96. doi : 10.1007 / s00253-004-1779-z . PMID  15630516 .
  2. Jump up^ Wolf, Fred R .; Nonomura, Arthur M .; Bassham, James A. (2004). “Growth and Branched Hydrocarbon Production in a Strain ofBotryococcus Braunii (Chlorophyta) 1″. Journal of Phycology . 21 (3): 388.doi : 10.1111 / j.0022-3646.1985.00388.x .
  3. ^ Jump up to:b Jian Qin (2005). “Bio-Hydrocarbons from Algae: Impacts of temperature, light and salinity on algae growth” (PDF) . Rural Industries Research and Development Corporation, Australia.
  4. Jump up^ Algal Oil Yields – Yield Data for Oil from Algae Strains, Algae Species with High Oil Yields. (2009-12-02). Retrieved on 2016-11-04.
  5. ^ Jump up to:b L.W. Hillen; et al. (1982). “Hydrocracking of the Oils of Botryococcus braunii to Transport Fuels” . Biotechnology and Bioengineering . 24 (1): 193-205. doi : 10.1002 / bit.260240116 . PMID  18546110 .
  6. ^ Jump up to:b J. Frenz; et al. (1989). “Hydrocarbon Recovery and Biocompatibility of Solvents for Extraction from Cultures of Botryococcus Braunii “. Biotechnology and Bioengineering . 34 (6): 755-62. doi : 10.1002 / bit.260340605 . PMID  18588162 .
  7. Jump up^ Biodiesel Production from Algae. US Department of Energy Aquatic Species Program
  8. Jump up^ Wolf, Fred R .; Nonomura, Arthur M .; Bassham, James A. (2004). “Growth and Branched Hydrocarbon Production in a Strain ofBotryococcus Braunii (Chlorophyta) 1″. Journal of Phycology . 21 (3): 388.doi : 10.1111 / j.0022-3646.1985.00388.x .
  9. Jump up^ “Fast, low energy, and continuous biofuel extraction from microalgae”. ScienceDaily . 2017-04-28.
  10. Jump up^ Why sequence Botryococcus braunii ? . Retrieved on 2016-11-04.
  11. Jump up^ Babu B, Wu JT (December 2008). “Production of Natural Butylated Hydroxytoluene as an Antioxidant by Freshwater Phytoplankton” (PDF) . Journal of Phycology . 44 (6): 1447-1454. doi : 10.1111 / j.1529-8817.2008.00596.x . PMID  27039859 .
  12. Jump up^ Nonomura, Arthur M. (May 5, 2006) “Methods and Compositions for Growth of Hydrocarbons inBotryococcussp.” US Patent 7,923,228
  13. Jump up^ “A new Japanese venture to pursue mass production of algae biofuel”. Shimbun Denki . 2011-07-12. Archived from the original on 2011-07-13.
  14. Jump up^ “Training of the Joint Venture by IHI and Neo-Morgan Laboratory for Bio-fuel production using Algae” . .