ISSN 1021-4437, Russian Journal of Plant Physiology, 2016, Vol. 63, No. 4, pp. 558–565. © Pleiades Publishing, Ltd., 2016.
Original Russian Text © S.S. Lapina, I.P. Maslova, V.P. Pchelkin, V.S. Bedbenov, K.S. Mironov, B.K. Zayadan, A.A. Userbaeva, M.A. Sinetova, D.A. Los, 2016, published in
Fiziologiya Rastenii, 2016, Vol. 63, No. 4, pp. 593–600.
Optimization of Prochlorothrix hollandica Cyanobacteria Culturing
for Obtaining Myristoleic Acid
S. S. Lapina
, I. P. Maslova
, V. P. Pchelkin
, V. S. Bedbenov
, K. S. Mironov
, B. K. Zayadan
A. A. Userbaeva
, M. A. Sinetova
, and D. A. Los
Institute of Plant Physiology, Russian Academy of Sciences, ul. Botanicheskaya 35, Moscow, 127276 Russia
Faculty of Biology and Biotechnology, Al-Farabi Kazakh National University,
pr. Al-Farabi 71, Almaty, Republic of Kazakhstan
Received November 20, 2015
Abstract—Plankton filament cyanobacteria Prochlorothrix hollandica is characterized by a very high content
fatty acids (FA) in the lipid membranes. Depending on culturing conditions of the cyanobac-
teria, total concentrations of myristic and myristoleic acids can reach 35% and those of palmitic and palmi-
toleic acids can reach 60% of all esterified FA cells. In P. h oll a ndica , a variety of monounsaturated FA is rep-
resented by myristoleic and palmitic acids, and by hexadecenoic (C
) acid with olefin bond of cis-configu-
ration, located in the Δ4 position. The process of intensive culturing for P. h o l l a n d i c a cells to yield a maximal
biomass in order to isolate the pure drug of myristoleic acid derivative has been optimized. The use of a three-
stage purification gives 30 mg of chromatographically pure myristoleic acid methyl ester from 17 g of P. ho l -
landica raw biomass (dry mass is 3 g), which is 1% of dry cell mass.
Keywords: Prochlorothrix hollandica, desaturase, fatty acids, lipids
Myristoleic acid (MA) is a monounsaturated fatty
acid (FA), 14:1Δ9 (sometimes designated as 14:1ω5).
MA induces apoptosis and necrosis in human prostate
cancer cells (LNCaP) in vivo with a minimum inhibi-
tory concentration of 9 μM. MA as a substrate is also
involved in the acylation of regulatory proteins, such as
tyrosine protein kinase Src is the main regulator in the
formation of osteoclasts, destructing bone tissue .
Cetyl myristoleate, a drug used for the treatment of
arthritis [2, 3] and fibromyalgia , is obtained from
esterification of free myristoleic acid with cetyl alcohol.
Typically, MA is isolated from the fruit of aquatic
grass staurogyne (Staurogyne repens) or from the seeds
or Myristicaceae plants; nutmeg, which is rich in
trimyristin—triglyceride with three saturated myristic
acid residues—and the African nutmeg (Pycnanthus
angolensis or P. ko m b o), from which oil (known as
combo oil) MA are obtained, are the most famous .
P. ko m b o seeds contain 56–61.6% of oil, which includes
58–64% of myristic acid and 19–26% of MA. The aver-
age trading price of MA in sales of leading manufactur-
ers of chemically pure reactants (Sigma–Aldrich, Cay-
man Chemical, MP Biochemicals, Protheragen, and
Wako Pure Chemical Industries) is approximately US
$1.000 per 1 g.
Photoautotrophic microalgae and cyanobacteria,
having a plastic metabolism and ability to adapt to a
variety of environmental conditions, are potential pro-
ducers of many useful compounds, including fatty
acids . The objectives of this work were the screen-
ing of cyanobacteria strains from microalgae collec-
tion of Institute of Plant Physiology Russian Academy
of Sciences (IPPRAS) for the search of potential MA
producer, the optimization of culturing conditions for
promising strains and the obtaining of chromato-
graphically pure MA methyl ester.
MATERIALS AND METHODS
The objects of the study were the strains of cyano-
bacteria from the collection of microalgae IPPRAS
(Moscow): Cyanobacterium sp. B-1200, Synechococ-
cus sp. B-266, Synechococcus sp. B-434, Synechocys-
tis sp. B-288, Thermosynechococcus vulcanus, Syn-
echococcus elongatus B-267, Synechococcus sp. B-468,
Phormidium laminosum B-407, Microcystis firma B-260;
and Prochlorothrix hollandica, which was originally
given by Prof. George S. Bullerjahn (Bowling Green
State University, Bowling Green, OH, United States).
The culturing conditions for Prochlorothrix hol-
landica and other cyanobacteria. Periodic culturing of
Abbreviations: MA—myristoleic acid; FAME—fatty acid methyl