Sys Rev Pharm 2020;11(7):89-97 A multifaceted review journal in the field of pharmacy Polyprenol and Dolichol Content in the Seed Tissues of Elaeis guineensis Jacq. from Commercial Seeds Mohammad Basyuni1*, Rahmah Hayati1, Ananda Ratu Tia1, Irma Deni1, Bejo Slamet1, Etti Sartina Siregar2 1*Department of Forestry, Faculty of Forestry, Universitas Sumatera Utara, Medan – Indonesia 2Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan – Indonesia Email for correspondence: [email protected] ABSTRACT Polyisoprenoid alcohols are found in the cells of all living organisms. This Keywords: Polyisoprenoid, polyprenol, dolichol, commercial seeds, non- work aims to investigate their distribution in the seed tissues of oil palms in commercial seed. commercial and non-commercial companies using two-dimensional thin layer chromatography (2D-TLC). The distribution of dolichols and polyprenols Correspondence: found in oil palm plants varies depending on the region. There is no difference Mohammad Basyuni1 in the distribution of polysoprenoids between Socfin Indonesia (Socfindo) CI 1*Department of Forestry, Faculty of Forestry, Universitas Sumatera Utara, Ganoderma susceptible, Socfindo Lame, and Indonesian Oil Palm Research Medan – Indonesia Institute (IOPRI) BSY2C seeds tissue (4.5 mg/g dry weight). The total Email: [email protected] polyprenol and dolichol in the seed tissues of oil palms from Socfindo MTG was higher than those from other oil palms: it was 6.1 mg/g dry weight and 4.7 mg/g dry weight, respectively. The highest polyisoprenoids concentration in the seed tissues of oil palms was found in those from Socfindo Yagambi. Polyprenol; longer chain lengths were observed in the oil palms of non- commercial (C75-C100) and dolichol of commercial seeds from IOPRI 05-73 (C70-C100). The seeds from Socfindo Yagambi were clearly separated from other samples, however, the seeds from commercial and non-commercial seeds (IOPRI B SMB and oil palm PT Harapan Sawita (OPHS) respectively were observed in one group. This study indicated that there are distinct groups of polyisoprenoid carbon chains randomly derived from cluster analysis in the origin of oil palm seeds. INTRODUCTION have been reported in oil palm leaf tissues as a Polyisoprenoid alcohols are found in the cells of all living chemotaxonomic marker [15]. Recently, we reported the organisms including bacteria, human, and plants. Polyprenols carbon chain pattern in E. guineensis polysioprenoids leaves are mainly present in plant photosynthetic tissues, whereas from different land uses, showing the yield gap in oil palm dolichols are present in plant roots [1][2]. Bioactive long- from planting materials [16]. The present study, therefore, chain isoprenoid alcohols (polyprenol) occur in plants. extends our work to investigate the polysoprenoids in oil Polyisoprenoid (polyprenol and dolichol) has been potent to palm seeds from the commercial and non-commercial anticancer and antibacterial activities [3]. Depending on the companies to obtain more insights distribution and source plants, the chain length of natural polyprenols varies occurrence of polyprenol and dolichol from oil palm. from six to forty isoprene units [4][5]. Polyprenols may play the role of scavengers for reactive oxygen and nitrogen MATERIALS AND METHODS species, protecting cells to oxidative stress. Un-saturated Plant Materials plant polyprenols and dolichols have the foremost Seventeen oil palm seed sources with triplicate were sampled antioxidant potential [6]. Polyprenols have been reported as in North Sumatra, Indonesia. These were collected as non- important active lipid components such as antivirus, commercial seeds from Universitas Sumatera Utara (USU), a hepatoprotective, and antitumor in pharmacology [7]. mangrove area 1% of salinity (MS1%), oil palm from Natural compounds classified as products of secondary smallholder in Marihat, Serdang Bedagai, North Sumatra metabolism are widely studied for their possible cellular (OPM), and private company of PT Harapan Sawita, functions of polyisoprenoids [8]. The seeds from Langkat, North Sumatra (OPTHS), whereas the oil palm monocotyledonous plants contained dolichol, a homologous seeds from Socfindo, Serdang Bedagai, North Sumatra varied of alpha-saturated polyisoprenoids alcohols, (private company), and Indonesian Oil Palm Research polyprenol, and a homologous series of alpha-unsaturated Institute (IOPRI), Medan, were categorized as commercial polyisoprenoid alcohols [9]. Dolichol synthesis established seeds as shown in Figure 1. The seeds were kept in an for the means asparagine-linked glycoproteins, such as average temperature at 25–27°C and the humidity was 83- tunicamycin in the establishment of GlcNAc-P-P-dolichol 84%. catalyzed by the enzyme dolichol phosphate N- acetylglucosamine-1-phosphate transferase [10][11]. The Chemical Compounds long-chain polyisoprenoid alcohols (polyprenols and A mixture of dolichol (C95-C110) standard compound with a dolichols), a cluster of compounds reported to develop in combination of polyprenol (C90-C100) was used to plants, indicates a major heterogeneity in terms of carbon- characterize the polyisoprenoids [1][17]. The silica gel 60 chain length [12]. TLC plates and reversed-phase of silica RP-18 HPTLC plates Oil palm plantation (Elaeis guineensis) is the main were obtained from Merck. commodity that supports the Indonesian economy and is well known as a profitable business [13][14]. According to our Polyisoprenoid Alcohols Isolation previous studies on the polyprenols of C45-C65 and C90- The protocol for the separation of polyisoprenoids was done C105 length chain, dolichols with C85-C105 chain-length as reported earlier [1][14]. The seeds of oil palm species 89 Systematic Reviews in Pharmacy Vol 11, Issue 7, July-Aug 2020 Basyuni et al. / Polyprenol and Dolichol Content in the Seed Tissues of Elaeis guineensis Jacq. from Commercial Seeds were kept in an oven at 60 °C for two days. The dried tissue used as chemotaxonomic marker [1][27]. Based on the (1–2 g) was pressed to powder and kept afloat in a 2/1 (v/v) molecular insights of polyprenol phosphates with diverse chloroform/methanol solution for two days. The extract of membrane-associated proteins in glycoconjugate, a cis/trans lipid was saponified at 65°C for 24 h in 50% ethanol with 2 geometry in polyprenol phosphate substrates of biosynthesis M KOH. The un-saponified lipids from a tissue sample were has been reported [28]. extracted into hexane, and the organic solution was The profile of polysoprenoids in non-commercial seeds is evaporated and dissolved again in hexane. The seeds described in Table 1. The total number of lipids in oil palm extracted (±100 µg) were used for each TLC plate. tissues was high in the USU region. It was 62.7 mg/d dry weight, whereas, in oil palms, from mangrove areas (1% of Two-Dimensional Thin Layer Chromatography (2D- salinity), the lipid value was 30.4 mg/d dry weight. The oil TLC) Analysis palm from a smallholder in Marihat (OPM) and PT Harapan 1D-TLC was formed 45 minutes earlier in a 20 cm × 3 cm Sawita (OPTHS) was 20.5 mg/d dry weight and 18.6 mg/d (silica gel) plate of glass in a soluble system of toluene-ethyl dry weight respectively. There was a difference in acetate (9:1) as was done previously [1]. The 2D-TLC polysoprenoids between the origins of oil palm crops; the reversed-phase C18 silica gel was finished with acetone as range value was 3.3 to 5.7 mg/g dry weight. The quantity of the soluble for 45 min. The state of the dilute polyisoprenoid polyprenol was highest in the oil palm from Marihat alcohols from the 2D-TLC was tough and envisaged with (smallholder) seeds tissue (3.5 mg/g) and the lowest in the iodine evaporate. To clarify the family to dolichols or ones from mangrove area (1% of salinity) (1.4 mg/g). polyprenols, a standard was applied to the line of sample for There was no difference in the dolichol content from USU the 1D-TLC. The occurrence of one family was verified in and OPTHS tissue of seeds palm; the value was 1.6 mg/g, 2D-TLC. The standard was used on the sample and delivered but the total dolichol content from the mangrove area was 2.0 with a solution system as previous reported. The mg/g, which is less than OPM, which is 2.2 mg/g. The chromatographic images were obtained and digitally scanned polysoprenoid in total lipid ranges from 1.9% to 2.2%. The with Canon G-2000 printer series. The polyisoprenoid group polyprenol from the oil palms within the USU area was was clarified by the correlation of movement on TLC in higher than the other oil palms, which was 1.8%. authentic dolichol or polyprenol standards using the 2- Furthermore, the dolicol of oil palm within the mangrove Dimensional phase. The polyprenols and dolichols found on area (1% of salinity) was higher than the other, which ranges the HPTLC plates RP-18 were measured with ImageJ ver. from 0.4% to 1%. Total polyprenol and dolycol in 1.46r [15] with dolichol and polyprenol standards. polysoprenoids content of oil palm from USU and OPM was the same value, it was 62% and 37%, respectively. In this Phylogenetic Analysis study, we found that the polyprenol and dolichol in the seed Phylogenetic analysis was carried out on a selected data of tissues of oil palm were on the same type within each non- 17 commercial and non-commercial seeds in palm oil from commercial origin and that it was type II distribution. the data of carbon-chain lengths of polyisoprenoids by log Table 3 shows the polyisoprenoid distribution in the oil (10) transformation by Euclidean distance [18]. palms of commercial seeds. The highest total lipid content in Dendrograms indicated the relationship by clustering seed tissues was found to be in Socfindo Lame species: 12.1 analysis used in the Un-Weighted Pair Group Method with mg/g dry weight, followed by Socfindo CI Ganoderma Arithmetic mean (UPGMA) MVSP ver.