April - June 2007 - Kasetsart University

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314 The crude hot water polysaccharide was partially purified by gel-filtration on Sepharose 6B column. Two fractions, SHP-F1 and SHP-F2, were collected (Figure 2). Attempts to completely separate the two fractions by decreasing flow rate from 1.2 to 0.8 ml/min was not successful. The curde hot water polysaccharide comprised of approximately 40% of fraction 1 (SHP-F1) and 60% of fraction 2 (SHP-F2). The fractions of SHP- F1 and SHP-F2 were repeatedly run using the same method at a lower flow rate of 0.5 ml/min. Absorbance (OD485) 4.5 4 3.5 3 2.5 2 1.5 1 0.5 SHP-F1 SHP-F2 0 0 10 20 30 40 50 60 70 80 Fraction number Figure 2 Elution profile of hot water polysaccharide by Sepharose 6B column chromatography. Absorbance (OD485) 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 0 10 20 30 40 50 60 70 80 Fraction number <strong>Kasetsart</strong> J. (Nat. Sci.) 41(2) A After the pool fraction of SHP-F1 was repeatedly applied on Sepharose 6B column, the purified SHP-F1 was obtained (Figure 3A). However, SHP-F2 still exhibited 2 peaks of absorbances, a small peak and a bigger one, designated as SHP-F2/1 and SHP-F2/2, respectively (Figure 3B). When the partially purified fractions of the hot water polysaccharide (SHP-F1 and SHP-F2) were subjected to cytotoxicity and anti HSV-1 assays, both fractions exerted non-toxicity on the growth of Vero cells at the maximum concentrations tested and had significantly higher anti HSV-1 activity than the crude hot water polysaccharide (about 4 and 2 times, respectively) (Table 1). The analysis of monosaccharide was performed by GC. It was found that SHP-F1 fraction contained only three sugars; rhamnose, ribose and arabinose, whereas, the SHP-F2 fraction contained rhamnose, ribose, arabinose, glucose, mannose, galactose and xylose. Both fractions contained rhamnose as the main sugar component (Table 2). Table 3 showed the comparison of proximate analysis of dried cells of Spirulina and the crude hot water extract polysaccharide. Results showed that dried cells consisted of 21.9% carbohydrate, 61.4% protein, 7.2% lipid and 7.2% Absorbance (OD485) 4.5 4 3.5 3 2.5 2 1.5 SHP- F2/ 2 1 0.5 SHP- F2/ 1 0 . 0 10 20 30 40 50 60 70 80 Fraction number Figure 3 Elution profile of SHP-F1 (A) and SHP-F2 (B) by Sepharose 6B column chromatography. B
ash. The crude hot water polysaccharide contained 42.5% carbohydrate, 31.0% protein, 12.9% ash and trace of calcium and sulfate. Table 4 shows the remaining carbohydrate and protein precipitated by various concentrations of trichloroacetic acid (TCA). These data suggested that at the highest concentration of TCA (50% TCA), 41% of the protein in dry cells was eradicated (a decrease from 31% to 18%), while the percentage of carbohydrate increased about 25% (from 42.5% to 53.1%). After <strong>Kasetsart</strong> J. (Nat. Sci.) 41(2) 315 the treated crude hot water polysaccharide (precipitated with 50% TCA) was tested for anti HSV-1 activity, results showed that the activity was not significantly different from the untreated crude hot water polysaccharide (data not shown). To determine the role of calcium ion and sulfate groups of the hot water polysaccharide in antiviral activity, calcium ion and sulfate groups in the polysaccharide were eliminated before testing for cytoxicity and anti HSV-1 activity. The results indicated that all of the calcium-free Table 1 Cytotoxicity and Anti HSV-1 activity of the hot water polysaccharide fractions from S. platensis. Fractions Cytotoxicitya Anti HSV-1b (IC50: µg/ml) (IC50: µg/ml) SHP-F1 > 50 5.25 SHP-F2 > 50 9.61 a maximum concentration of compound for cytotoxicity test was 50 µg/ml compound was non-toxic to the growth of Vero cells when IC50 >50 µg/ml (if compound was toxic on the growth of Vero cells, the compound will be subjected to the serial dilution for determination of IC50 value) b % inhibition of HSV-1; < 25%= inactive, 25-35%= weakly active, >35-50% = moderately active, > 50%= active (the compound will be subjected to the serial dilution for determination of IC 50 value) Table 2 Sugar composition of fractions of the hot water extract polysaccharide. Fractions % Sugar composition Rhamnose Ribose Arabinose Glucose Mannose Galactose Xylose SHP-F1 75.6 13.4 11.0 - - - - SHP-F2 30.4 27.1 10.0 18.2 7.5 4.5 2.3 Table 3 Composition of S. platensis powder and crude hot water polysaccharide. Composition Dry weight (%) Spirulina powder Crude hot water polysaccharide Carbohydrate 21.9 ± 0.8 42.5 ± 0.3 Protein 61.4 ± 1.1 31.0 ± 0.8 Lipid 7.2 ± 1.3 0 Calcium -* 0.123 ± 0.0006 Sulfate -* 1.44 ± 0.03 Ash Mean ± standard deviation (n = 3) 7.2 ± 0.1 12.9 ± 0.4 * It was not determined
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April - June 2007 Volume 41 Number
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KASETSART JOURNAL NATURAL SCIENCE T
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Kasetsart J. (Nat. Sci.) 41 : 205 -
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harvesting time which started from
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y Chen and Paull (2000). Figure 1 a
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pineapple fruit allowed the accumul
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Kasetsart J. (Nat. Sci.) 41(2) 215
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Cluster analysis Cluster analysis u
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Table 3 (Continued) 1 2 3 4 5 6 7 8
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CONCLUSION AFLP markers classified
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difference of soil texture used in
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under wet soil condition planting.
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tropics. Following the expansion, w
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sealed in a plastic box with 1 cm w
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moisture content increment after so
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Frequency Frequency 30 20 10 0 20.0
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School, Kasetsart University. The a
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for combining ability of lines (Lam
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selective mass sibbing within indiv
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Although most of S 2-interfamily hy
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composite sets as used in this stud
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days. The numbers of calli producin
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the report of Ching (1982) showing
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clearly amplified bands generated b
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caused by either the recombination
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Kuhlmann, V. and B. Foroughi-Wehr.
Page 63 and 64: Xanthomonas fuscans subsp. aurantif
Page 65 and 66: Kasetsart J. (Nat. Sci.) 41(2) 265
Page 67 and 68: was collected and washed with 70% e
Page 69 and 70: expected size was amplified with 35
Page 71 and 72: eaction technique has been used for
Page 73 and 74: Schaad, N.W., D. Opgenorth and P. G
Page 75 and 76: MATERIALS AND METHODS Model descrip
Page 79 and 80: calculated TF value calculated TF v
Page 81 and 82: sincere gratitude and deep apprecia
Page 83 and 84: 1989). In monogastrics, the inclusi
Page 85 and 86: of all LLS samples in this study we
Page 87 and 88: Melbourne, Parkville, Victoria. Goe
Page 89 and 90: considered responsible for low prod
Page 93 and 94: mineral supplementation suggested b
Page 95 and 96: Body weight chane (kg/head) 32 30 2
Page 97 and 98: CONCLUSION AND RECOMMENDATION With
Page 99 and 100: SAS. (Statistical Analysis System).
Page 101 and 102: genes covering a variety of desirab
Page 103 and 104: mg/l NAA and 0.5 mg/l zeatin) incub
Page 105 and 106: 5. Purification by various sucrose
Page 107 and 108: as the culture period increased. So
Page 109 and 110: culture, pp. 1-20. In L.C. Fowke an
Page 111 and 112: Kasetsart J. (Nat. Sci.) 41 : 311 -
Page 113: GC. Analytical methods Total carboh
Page 117 and 118: spirulan (H-SP), and a desulfated c
Page 121 and 122: cells often displayed an increased
Page 123 and 124: LITERATURE CITED Bell, T.A. and D.V
Page 125 and 126: for alternative sources of supply.
Page 127 and 128: BR2.1.2 formed the same clade with
Page 129 and 130: supplement with glutamate (Iida et
Page 131 and 132: optimal for S. limacinum BR2.1.2 fo
Page 133 and 134: fluorescent lamp at 1.5 kLux develo
Page 137 and 138: Wisconsin, USA). Total extracted RN
Page 139 and 140: RESULTS Cloning and sequencing of m
Page 141 and 142: Expression of rmIL-2 and purificati
Page 143 and 144: other strains of mice have differen
Page 145 and 146: dose interleukin-2 therapy promotes
Page 147 and 148: The chitosano-oligosaccharides are
Page 149 and 150: enzyme and cells were maximized by
Page 151 and 152: of crude chitosanases. Fortunately,
Page 153 and 154: Relative activity (%) 100 80 60 40
Page 155 and 156: Uchida, K. Tateishi, O. Shida and K
Page 157 and 158: MATERIALS AND METHODS 1. Materials
Page 159 and 160: without coating and coated with pec
Page 161 and 162: in Table 5-7. It was found that the
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the cultures at 10,200 × g for 15
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incubation and reached the final pH
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y lysine- and tyrosine-decarboxylas
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during the stages of ripening as su
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As the 25 companies were divided in
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All the data comes up with informat
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as piece “A” or straight as pie
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et al. (1998); Zeng (2000); and Tal
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p 1, i * Kasetsart J. (Nat. Sci.) 4
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c ≤ c2 jq2 j n q p k p * 2, j , ,
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algorithm would consider RM 5, prio
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the maximum deviation of about 10%.
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Stock Quantities Using Heuristic Op
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which dynamically and automatically
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2. Framework architecture and compo
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Risk (VaR) calculation which was im
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Throughput (job/sec) 7 6 5 4 3 2 1
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Speed up 40 35 30 25 20 15 10 5 0 F
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and discovery, resource selection a
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April - June 2007 - Kasetsart University

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