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Production of Xylanase by Recombinant Bacillus subtilis DB104 Cultivated in Agroindustrial Waste Medium
2016
Helianti, Is | Ulfah, Maria | Nurhayati, Niknik | Suhendar, Dadang | Finalissari, Anita Kusuma | Wardani, Agustin Krisna
A recombinant Bacillus subtilis DB104 strain harbouring recombinant plasmid pSKE194 containing an Open Reading Frame (ORF) of endoxylanase and its indigenous promoter from the wild-type B. subtilis AQ1 strain was constructed. This recombinant B. subtilis DB104 strain had higher endoxylanase activity than the nonrecombinant B. subtilis DB104 strain in standard media, such as Luria Bertani (LB) and LB with xylan. The agroindustrial wastes corncobs and tofu liquid waste were chosen as cost-effective carbon and nitrogen sources, respectively, to test the economics of xylanase production using the recombinant B. subtilis DB104 at a larger scale. Submerged fermentation using a 4.5 L working volume fermentor with tofu liquid waste and 4% corncobs produced maximum xylanase activity of 1296 ± 1.2 U/mg (601.7 ± 0.6 U/mL) after 48-hour fermentation at 37°C with 150 rpm agitation; this is more than twofold higher than the activity produced in an Erlenmeyer flask. This is the first report of high xylanase activity produced from recombinant B. subtilis using inexpensive medium. During fermentation, the xylanase degrades corncobs into xylooligosaccharides, showing its potential as an enzyme feed additive or in xylooligosaccharide production.
Show more [+] Less [-]Expression of Recombinant Sugarcane Streak Mosaic Virus Coat Protein Gene in Escherichia coli
2016
Hamdayanty, Hamdayanty | Hidayat, Sri Hendrastuti | Damayanti, Tri Asmira
Sugarcane streak mosaic virus (SCSMV) is an important virus causing mosaic disease in sugarcane and transmitted through the cutting cane. Commercial antiserum to detect SCSMV and to monitor the disease development is not available. The research was conducted to produce antigen of SCSMV coat protein (SCSMV-CP) through overexpressing it on bacterial expression which will be used for antiserum production. SCSMV-CP was amplified using specific primers for CP gene containing BamHI and HindIII restriction enzyme sites and cloned into pTZ57R/T. Subsequently, the SCSMV-CP was subcloned into pET28a and transformed on Escherichia coli BL21(DE3) and Rosetta-gami(DE3)pLysS. The concentration of isopropyl β-d-thiogalactopyranoside (IPTG), incubation temperature, and bacterial harvesting time after IPTG induction were optimized. SCSMV-CP gene was successfully amplified with size ±855 bp, subcloned into vector expression, and expressed in insoluble fraction either in both bacterial host. Optimal protein expression of SCSMV-CP recombinant was obtained at 25°C with IPTG concentration 0.25–1.00 mM and harvested at 9–12 hours after IPTG induction in E. coli BL21(DE3), and at 30°C with IPTG concentration 0.25–1.00 mM and harvested 3–12 hours after IPTG induction in E. coli Rosetta(DE3)pLysS. SDS-PAGE analysis showed that protein size of SCSMV-CP recombinant was ±35.4 kDa.
Show more [+] Less [-]Growth, Morphology and Growth Related Hormone Level in Kappaphycus alvarezii Produced by Mass Selection in Gorontalo Waters, Indonesia
2016
Fadilah, Siti | Alimuddin, | Pong-Masak, Petrus Rani | Santoso, Joko | Parenrengi, Andi
The use of high quality seed can support the success of the seaweed cultivation. This study was conducted to evaluate the growth performance, morphology and growth related hormone level of brown strain seaweed Kappaphycus alvarezii seed produced by mass selection. Selection was performed in the Tomini Gulf, Gorontalo, based on mass selection of seaweed seed protocol with a slight modification in cut-off 10% of the highest daily growth rate. Selection was carried out for four generations. The selected 4th generation of seed was then used in cultivation performance test in the Celebes Sea, North Gorontalo, for three production cycles. The results showed that the selected K. alvarezii has higher clump weight and daily growth rate, longer thallus, more number of branches, and shorter internodes compared to the unselected control and seaweed from the farmer as external control. Furthermore, total sugar content, levels of kinetin hormone and kinetin:indole-3-acetic acid ratio were higher in selected seaweeds than that of unselected control and external control. Thus, mass selection method could be used to produce high growth of seed, and kinetin and indole-3-acetic acid play an important role in growth of K. alvarezii.
Show more [+] Less [-]Marine Microalgae Tetraselmis suecica as Flocculant Agent of Bio-flocculation Method
2016
Kawaroe, Mujizat | Prartono, Tri | Sunuddin, Adriani | Saputra, Deni
Microalgae harvesting is an important part in microalgae cultivation system. Techniques for harvesting marine microalgae which are commonly used are centrifugation, filtration and flocculation. These techniques still have some disadvantages, such as not environment friendly, and high usage of energy and cost. Bio-flocculation harvesting technique using microalgae as a flocculant agent can be an alternative way to solve these problems. In this research, mixing of Tetraselmis suecica (flocculant) with Chlorella sp. and Nannochloropsis sp. (non-flocculant) in ratios of 1:4, 2:4, 3:4 and 4:4 (v/v) has been conducted to obtain percent recovery of marine microalgae harvest. The results showed that T. suecica as flocculant agent can fasten the harvesting of Chlorella sp. and Nannochloropsis sp. It was shown by the increase of percent recovery value of Chlorella sp. from 51.14 ± 1.07% to 67.34 ± 0.67% and Nannochloropsis sp. from 20.52 ± 1.17% to 42.43 ± 0.40% during the first hour of flocculating process. Our result showed that bio-flocculation is an environment friendly technique which can be applied to harvest marine microalgae.
Show more [+] Less [-]Diversity and Abundance of Carabidae and Staphylinidae (Insecta: Coleoptera) in Four Montane Habitat Types on Mt. Bawakaraeng, South Sulawesi
2016
Qodri, Agmal | Raffiudin, Rika | Noerdjito, Woro Anggraitoningsih
Carabidae and Staphylinidae are the two beetle families frequently found to be most abundant and diverse in forest ecosystem. Their roles especially as generalist predators are important in forest ecosystem. No studies reported their diversity and abundance in forest ecosystem on Mt. Bawakaraeng, specifically in montane habitat yet. This study was aimed to analyze diversity and abundance of Carabidae and Staphylinidae in four montane habitat types, i.e. agricultural area, pine forest, eucalypts and natural forest (1,835 m asl), and natural forest (2,165 m asl). They were collected using pitfall traps. A total of 42 carabid beetles belonging to nine species and 260 staphylinid beetles belonging to 37 species were collected. Diversity and abundance of Staphylinidae were higher than Carabidae, this is predicted because of higher mobility in Staphylinidae compared to Carabidae. In Carabidae, the highest species richness was recorded in agricultural area, whereas the highest species richness of Staphylinidae occurred in natural forest (2165 m asl). In Staphylinidae, the two largest subfamilies are Aleocharinae and Staphylininae. Aephinidius adelioides occupied the highest abundance of Carabidae and found in agricultural area. The differences in each montane habitat type are presumed to cause variation in species richness of soil beetles, especially for Carabidae.
Show more [+] Less [-]Alteration of Leaf Anatomy of Handeuleum (Graptophyllum pictum L. Griff) due to Gamma Irradiation
2016
The leaves of the plant handeuleum (Graptophyllum pictum L. Griff) have long been used for traditional medicine in several regions in Indonesia. This study was aimed to determine the effect of gamma irradiation rate on the anatomy and phytochemical content of the leaf. The rates of gamma rays used were 0, 15, 30, 45, 60, 75, 90, and 105 Gy. Our results showed that gamma ray irradiation rate of 30 Gy produced leaves that contain anthocyanins and carotenoids, with the highest number of stomata and stomatal density compared with control plants. Stomatal index was found highest in the leaves with 45 Gy of gamma irradiation. High-rate gamma ray irradiation produced rigid, thick, and frangible leaves. A high rate of gamma irradiation, i.e. 75, 90, and 105 Gy, produces bigger palisade, sponges, and upper epidermis than the control plants, respectively. Our results showed an association between increasing rate of irradiation with alterations in the structure of leaf anatomy and phytochemical content of handeuleum.
Show more [+] Less [-]Diversity and Abundance of Cerambycid Beetles in the Four Major Land-use Types Found in Jambi Province, Indonesia
2016
Fahri, | Atmowidi, Tri | Noerdjito, Woro Anggraitoningsih
Longhorn beetles (Coleoptera: Cerambycidae) have an important function in the ecosystem, i.e. bioindicators, saproxylic, pollinators, and as food of other organisms. Land cover changes due to land use can disrupt the natural balance of the ecosystem, which can result in a decrease of cerambycid diversity. Cerambycid species diversity was evaluated in four land types, i.e. jungle-rubber, rubber plantations, oil palm plantations, and felled jungle-rubber. Collections of cerambycid beetles were conducted by using artocarpus trap, made by freshly cut Artocarpus heterophyllus branches. Collections of beetles were made on day 4th, 7th, 10th, 13th, and 16th after the traps were set up. In the four land-use types in Jambi province, we collected 72 species including 34 morphospecies of cerambycids, consisting of 42 species from the jungle-rubber, 39 species from rubber plantations, 16 species from oil palm plantations, and 28 species from felled jungle-rubber. Cerambycid diversity was highest in jungle-rubber (H'=3.23), followed by rubber plantation (H'=2.67), felled jungle-rubber (H'=2.38), and oil palm plantations (H'=2.01). Highest similarities of cerambycid communities occurred in the rubber plantation–felled jungle-rubber (51.2), followed by jungle-rubber–rubber plantation (50.0), rubber plantations–oil palm plantations (43.5), oil palm plantation–felled jungle-rubber (42.4), jungle-rubber–oil palm plantations (35.3), and jungle-rubber–felled jungle rubber (34.8). The number of cerambycid species and individuals collected was highest on day 7th.
Show more [+] Less [-]Molecular Phylogenetic Screening of Withania somnifera Relative From Indonesia Based on Internal Transcribed Spacer Region
2016
Hidayat, Topik | Priyandoko, Didik | Wardiny, Putri Yunitha | Islami, Dina Karina
Withania somnifera (family Solanaceae), known commonly as Ashwaganda, is one of the important medicinal plants, and recent studies reported that Withanone, one of the chemical components in this plant, has ability to kill cancer cell. Because of endemic state of this plant to South Asia, exploring plant species under the same family which grow well in Indonesia has been of interest. The purpose of this study was to screen the Indonesian plant which has strong phylogenetic relationship with Ashwaganda. Thus, phylogenetic analysis using DNA sequences of internal transcribed spacer (ITS) region was conducted. Thus, 19 species of Solanaceae and two species of Convolvulaceae as outgroup were examined. Five ITS regions of Ashwaganda retrieved from GenBank were included in the phylogenetic analysis. Parsimony analysis showed that Indonesia Solanaceae comprises seven groups which is consistent with the global Solanaceae relationship as previously reported. Furthermore, our study revealed that two species, Physalis angulata and Physalis peruviana, are relative to W. somnifera. Morphologically, they share characters of flower and fruit. This result indicated that these two species are potential to have similar chemical properties as Ashwaganda, thus we can have new variants of Withanone originated from Indonesia with similar effect.
Show more [+] Less [-]Two Newly Recorded Species of the Lobster Family Scyllaridae (Thenus indicus and Scyllarides haanii) From South of Java, Indonesia
2016
Wardiatno, Yusli | Hakim, Agus Alim | Mashar, Ali | Butet, Nurlisa Alias | Adrianto, Luky
Two species of slipper lobster, Thenus indicus Leach, 1815, and Scyllarides haanii De Haan, 1841, are reported for the first time from the coastal waters of South of Java, part of the Indian Ocean. A total of two specimens, one specimen of T. indicus from Palabuhanratu Bay and one specimen of S. haanii from Yogyakarta coastal waters, were collected in April and September 2015, respectively. Descriptions and illustrations of the morphological characteristics of the two species and their habitat are presented.
Show more [+] Less [-]Identification and Behavioral Evaluation of Sex Pheromone in Xanthopimpla pedator (Fabricius)—A Serious Pupal Parasitoid of Tropical Tasar Silkworm Anthereae mylitta Drury
2016
Lakshmi Marepally | Gaddam Benarjee
Xanthopimpla is a major parasitoid of silk worm cocoons. The female Xanthopimpla pedator (Fabricius) lays the eggs in male cocoons. Control of this infestation with pesticides is not recommended because of its concealed behavior. Various control methods were found to be inefficient. Ecofriendly management is the best strategy that can be applied. We have studied the sex communication in Xanthopimpla pedator (Fabricius), which helps to develop management strategy. Bioassays were done in the laboratory by using olfactometer and pheromone extraction chambers. It was found that female Xanthopimpla produces sex pheromones. The results show a strong attraction of male by female Xanthopimpla. Present results with male and female volatiles also show that female volatiles attract male Xanthopimpla. Fractionation of female volatiles by column chromatography has proven that 20% fraction has highest attraction of males by females.
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