Sugar Cane Extract Research Articles

Expression of fructokinase isoforms in the sugarcane culm

Two isoforms of fructokinase (FRK), FRK1 and FRK2, are present in sugarcane ( Saccharum spp. var N19) internodal tissue. Both isoforms are highly specific for fructose as the hexose substrate. FRK2 is inhibited by fructose concentrations exceeding 0.1 mM and 50% inhibition is attained at 230 μM ( K i (Fru) = 160 μM), while FRK1 activity is not negatively affected even at 1.0 mM fructose. The ratio of FRK2 to FRK1 activity is dependent on the developmental stage of the tissue. FRK1 appears to be the isoform that is preferentially expressed in mature tissue. Total FRK activity decreases during tissue maturation. This is the result of changes in expression of the isoforms and not inactivation of existing protein. A mathematical method that allows accurate estimation of the activities of the two isoforms of FRK in crude sugarcane extracts is presented.

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens.

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.

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Molecular characterization of Fiji disease fijivirus genome segment 9.

This is the first report of sequence from Fiji disease fijivirus (FDV), the type member of the genus Fijivirus of the family Reoviridae. FDV genome segment (S9) comprised 1843 nt and contained two non-overlapping ORFs, separated by a 57 nt intergenic region. S9 ORF 1 comprised 1008 nt and encoded a 335-amino-acid polypeptide (predicted molecular mass 38.6 kDa), while ORF 2 comprised 627 nt and encoded a 208-amino-acid polypeptide (predicted molecular mass 23.8 kDa). The 5' and 3' non-coding regions were 49 and 102 nt, respectively. The S9 terminal sequences were 5' AAGUUUUU------UGUC 3', and located immediately adjacent to these sequences were 12 bp imperfect inverted repeats. The entire S9 ORF 1 and the hydrophilic regions of S9 ORF 2 were each expressed as a fusion protein with the maltose-binding protein in Escherichia coli. Antibodies produced against the ORF 1 fusion protein reacted strongly with a protein of approximately 39 kDa present in both crude extracts of FDV-infected sugarcane and partially purified FDV preparations. In contrast, antibodies raised against the modified ORF 2 fusion protein did not react with any proteins in the same samples. Further, polyclonal antibodies produced against partially purified FDV reacted with the ORF 1, but not the modified ORF 2, fusion protein. These results indicate that FDV S9 ORF 1 encodes a major structural protein, while ORF 2 probably encodes a non-structural protein.

Sugar-cane juice induces pectin lyase and polygalacturonase in Penicillium griseoroseum

The use of other inducers as substitutes for pectin was studied aiming to reduce the production costs of pectic enzymes. The effects of sugar-cane juice on the production of pectin lyase (PL) and polygalacturonase (PG) by Penicillium griseoroseum were investigated. The fungus was cultured in a mineral medium (pH 6.3) in a rotary shaker (150 rpm) for 48 h at 25oC. Culture media were supplemented with yeast extract and sucrose or sugar-cane juice. Sugar-cane juice added singly to the medium promoted higher PL activity and mycelial dry weight when compared to pectin and the use of sugar-cane juice and yeast extract yielded levels of PG activity that were similar to those obtained with sucrose-yeast extract or pectin. The results indicated that, even at low concentrations, sugar-cane juice was capable of inducing pectin lyase and polygalacturonase with no cellulase activity in P. griseoroseum.

Production of ?-galactosidase by Monascus grown on soybean and sugarcane wastes

Extracts of both sugarcane and soybean wastes supported the growth of Monascus but sugarcane waste was superior for the production of α-galactosidase. An aqueous extract prepared from 5% (w/v) soybean waste and 7% (w/v) sugarcane waste gave the best result and was superior to the standard peptone/glucose/yeast extract medium. Liquid-solid mixtures were slightly less effective. Enzyme production could be enhanced by adding raffinose. Enzymatic hydrolysis of p-nitrophenyl-α-D-galactoside was optimal at pH 4.5. Raffinose and stachyose were hydrolysed to sucrose and galactose.

Growth and sporulation of Ustilago scitaminea, in vivo and in vitro

Two hyphal forms of Ustilago scitaminea were observed in vivo: dikaryotic vegetative hyphae in the sugarcane stems and monokaryotic sporogenous hyphae in the peripheral layers of the whip, a sorus-bearing structure on infected stalks. The fungus was easily isolated from sugarcane tissue and cultured on six different media. A brown cerebriform colony was usually the first form to grow from tissue. A white mycelial colony commonly arose from the brown colony after 1–2 wk and grew rapidly over its surface. On a medium containing extracts of sugarcane, the fungus developed into black colonies in which teliospores were formed. Teliospore production was enhanced and development of black colonies prolonged if covered by a thin film of liquid medium. Without a moist covering, black colonies were commonly overgrown by white mycelium.

The effect of sugarcane extracts on the growth of the pathogenic fungus, Colletotrichum falcatum

A red pigment which is induced in sugarcane stalks ( Saccharum officinarum L.) upon infection with Colletotrichum falcatum Went was assayed for its effect on spore germination and radial mycelial growth of C. falcatum. The pigmented extracts from infected plants inhibited both spore germination and radial mycelial growth. The inhibition caused by the pigment was modulated by the presence of growth stimulating factors in the sugarcane extracts. Partial characterization of the crude pigment showed the presence of luteolinidin, luteolin, chlorogenic acid and a glucoside of luteolinidin. While in vitro spore germination was inhibited by the pigment, spores exposed to extracts from uninfected sugarcane stalks had nearly completed germination within 14 h. The first pigment component in infected tissue did not appear until at least 24 h after infection, indicating that the pigment components may function to slow the growth and spread of the mycelia in vivo but may not play a role in controlling spore germination in vivo. The concentration of phenolic compounds in the cane tissue could not be correlated with resistance to C. falcatum.

A survey of the presence of glutamate synthase in plant cell suspension cultures

Evidence was sought for the presence of glutamate synthase (EC 2.6.1.53) in extracts from suspension cultures of six higher plant species not previously examined. The level of glutamate synthase measured was above the level of glutamic dehydrogenase (EC 1.4.1.2, 1.4.1.4) in extracts of soybean, parsley, okra, and cotton. Glutamate synthase was detectable but less than glutamic dehydrogenase in extracts of sugarcane. Glutamate synthase was not detected in extracts of peanut. Evidence for two glutamate synthases, each specific for one pyridine nucleotide, was obtained with cultures of carrot. Glutamate synthase has now been detected in eight and possibly nine species representing four and possibly five families of higher plants.

Biosynthesis of sucrose phosphate with sugar cane leaf chloroplasts.

Leloir and his collaborators (7, 11, 12) and others (1, 3, 6, 9, 13, 15, 16) have shown that synthesis of sucrose in extracts of a number of plants can occur by 2 different enzymic reactions: A) UDP3-Dglucose + D-fructose sucrose + UDP; B) UDPD-gltucose + D-fructose 6-P = sucrose-P + UDP. In reaction (A) D-fructose serves as the glucosyl acceptor and the reaction is freely reversible. In reaction (B) in which D-fructose 6-P is the acceptor, the sucrose-P formed is hydrolyzed by a phosphatase in a reaction which is practically irreversible, producing free sucrose. It is of interest to note that small amounts of sucrose-P have been detected among label-d photosynthetic products in plants (2, 5). In a previous communication Frydman and Hassid (8) showed that extracts of sugarcane leaves contained an enzyme which produced sucrose from UDPD-glucose and D-fructose, but neither sucrose nor sucrose-P was formed when D-fructose wvas reDlaced bv D-fructose 6-P in the reaction mixture. This indicated that only the enzyme which catalyzed reaction (A) was present in the extract. However, when chloroplasts isolated from the sugarcane leaves were used -as the enzyme source, sucrose was formed from UDP-D-glucose and D-fructose 6-P as well as from UDP-D-glucose and D-fructose. Since sucrose-P was not detected in the incubation mixture containing UDP-D-glucose and D-fructose 6-P, it could not be decided whether the sucrose was formed by enzymic removal of P from sucrose-P or by glucosylation of D-fructose produced by the action of a phosphatase onl D-fructose 6-P. Bird and Stocking (4) have demonstrated the synthesis of suicrose from UDP-D-glucose and D-fructose or D-fructose 6-P by chloroplasts isolated in a nonaqueous medium; but they were also unable to detect the formation of sucrose-P. Evidence is presented in this paper that the chloroplasts of sugarcane leaves and spinach leaves contain, in addition to the enzyme that forms sucrose from UDP-D-glucose and D-fructose, the enzyme that catalyzes the formation of sucrose-P from UDP-Dglucose and D-fructose 6-P.