Effect of phenylalanine in cell suspension cultures of M. oleifera Lam.

3.4. Effect of phenylalanine in cell suspension cultures of M. oleifera Lam.

Results of one-way ANOVA test, p 0.010 (p 0.05) showed that the phenylalanine concentration variations affect the total content of phenolic acids. Calculation of the

total concentration of phenol acid using a regression equation tannic acid standard curve y = 4,091x-0,001. In the cell suspension cultures of M. oleifera precursor feeding was used phenylalanine at concentrations of 0 ppm (control), 5 ppm, 10 ppm and 15 ppm.

Table 2. Total Acid Phenol concentration and celldry weightof M. oleifera In Treatment of Precursor Feeding Phenylalanine

Treatment

Total phenolic acid (ppm)

Cell Dry wieght (mg)

0 ppm a 0,63 ± 0,15 6,9 ± 0,80

5 ppm b 1,60 ± 0,37 7,3 ± 0,61

10 ppm

1,26 ab ± 0,31

7,8 ± 0,72

9,7 ± 1,46 Based on Table 2, it is known that precursor feeding of phenylalanine treatment of 0-

15 ppm

1,52 b ± 0,29

15 ppm concentration showed an increase in dry weight and total phenolic acids compared to control. The highest dry weight contained in the treatment of precursor feeding phenylalanine concentration of 15 ppm is 9.7 ± 1.46 mg and the lowest concentration found in the control 0 ppm was 6.9 ± 0.80 mg. The highest concentration of phenols contained in the treated precursor feeding phenylalanine concentration of 5 ppm was 1.60 ± 0.37 ppm and the lowest concentration found in the control 0 ppm was 0.63 ± 0.15 ppm. Increased cell dry weight due to the addition of exogenous amino acids such as phenylalanine. This is consistent with the statement [34] which states the addition of phenylalanine in the cell suspension culture medium can increase the content of endogenous phenylalanine. So that the excess amount of carbon that can be used for the process of growth [35].

Increased total phenol acid because phenylalanine is the main precursor in the biosynthesis of phenolic compounds. So the more phenylalanine is absorbed the more the total phenolic compounds are produced. This is in line with the statement [36] that the addition of phenylalanine affect the content of phenolic compounds as total acid phenylalanine is the main precursor in the metabolic process through phenylpropanoid, thus increasing the amount of phenylalanine which will be absorbed increase income PAL enzymes that affect the process of formation of

Sept em ber 8 th – 9 th 2015, Facult y of Biot echnology – Universit as At m a Jaya Yogyakart a

1.08 mg/g compared to the control that was 6.24 ± 0.68 mg/g wet weight and increase callus 0, 30 grams compared to the control that was 0.26 grams.

Phenylalanine concentrations of 5 and 15 ppm significant effect on total acid production process phenol compound in cell suspension cultures of M. oleifera compared with controls. Improvement of secondary metabolites such as phenolic acid compounds can be due to an increase in PAL enzyme which included the key enzymes before entering cinnamat acid pathway [19]. In the treatment of phenylalanine concentration of 10 ppm decreased phenolic compounds while increasing cell dry weight. This is because in the process of cell suspension cultures, callus M. oleifera not completely dispersed so that the cell surface area in contact with the medium are also smaller. This indicates that cell size affects the level of absorption of nutrients in the growth medium. By [37] states that the cell size has an influence on the level of absorption of nutrients, which the smaller cell size has the ability to absorb nutrients faster than larger cells. It has been proved by [37] to study the morphology and size variations of phytoplankton cells and concluded that the level of sinking or absorption of good nutrition is limited to the size of 20 m

cells.