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Wijanarka, W
Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro

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Produksi Enzim Inulinase Khamir Pichia manshurica DUCC Y-015 Dari Tepung Umbi Dahlia (Dahlia variabilis Willd.) Dengan Variasi Konsentrasi Magnesium Sulfat (MgSO4.7H2O) Dan Waktu Inkubasi

Jurnal Akademika Biologi Vol. 4 No. 2 April 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Dahlia tubers (Dahlia variabilis Willd.) contain inulin which can be hydrolyzed by the inulinase enzyme (E.C.3.2.1.7) into fructose monomer units. Application of  inulinase enzyme is used in the production of HFS (High   Fructose  Syrup)  dan  FOS  (Fructo-oligosaccharides).  Inulinase  can  be  produced  by  several microorganisms  including  inulinolytic  yeast  Pichia  manshurica  DUCC  Y-015.  One  of  the  factors  that influence the production of enzyme inulinase is macro minerals and incubation time on production medium. This study aims to determine the concentration of magnesium sulphate (MgSO4.7H2O) and the most optimal incubation time in producing the inulinase  enzyme. The research was carried out experimentally using a Randomized Block Design factorial. The first factor is the concentration MgSO4.7H2O those are 0,5 g/L; 1 g/L; and 1,5 g/L. The second factor is the variation of the incubation time, those are 12 hours; 18 hours; and24 hours, repetition was performed three times. Data were analyzed using ANOVA with 5% significant level(α = 0,05) and Duncan Test for further analysis. The results showed that the variation of the concentration ofMgSO4.7H2O has not been able to increase the production of inulinase enzyme, while the incubation time of18 hours produced the inulinase enzyme activity of 0,9605 IU/mL. Keywords:  Inulinase,  Dahlia  variabilis  Willd.,  Pichia  manshurica  DUCC  Y-015,  MgSO4, Incubation Time

Produksi Enzim Protease Aspergillus Flavus Pam-25 Dengan Variasi Ph Dan Waktu Inkubasi

Jurnal Akademika Biologi Vol. 4 No. 2 April 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Enzyme is a biokatalisator which can increase the speed of the reaction without join react. Protease enzyme is one of the enzymes that have a high economic value because its application is extensive in the field of industry. Protease is an enzyme that catalyzes the proteolytic peptide bonds in proteins termination. This research aims to determine the effect of pH and incubation time on the production of protease enzyme from A. flavus PaM-25.. This research aims to know the influence of the variation of the pH of the incubation time and against the production of the enzyme protease of A. flavus PaM-25. The research  was conducted at the laboratories of Microbiology Department of Biology, Faculty of Science and Mathematics, University of Diponegoro. Variables observed were protease activity, protein content and specific activity. Research using randomized complete block design (RAK) with two factors. The first factor is variation of pH, P1 (pH 7), P2 (pH 8), and P3 (pH 9), while the second factor is the incubation time T5 (5th day), T6 (6th day) and T7 (7th day) with repeated 3 times. Research data analyzed by Analysis of variance (ANOVA) and continued by Least Significant Difference test and Duncan Significant Difference Test at test level 5%. The results showed that the Aspergillus flavus PaM-25 has the capability of producing alkaline protease enzymes with pH range 7.0-9.0. The highest activity of proteases of   A. flavus PaM-25 retrieved on 7th day incubation time of treatment with protease activity value 1.94 U/mL, while pH treatment has no effect. The highest levels of protein found in the treatment of pH 7 and 5th days of incubation time of 1.05 mg/mL. The value of the highest purity of enzymes found in the combination of treatment pH 9 and 7th day incubation time  of 12.91 U/mg protein

TRANSFORMASI DAN KLONING PLASMID PJ804:77539 PADA E.coli TOP’10

Jurnal Akademika Biologi Vol. 6 No. 1 Januari 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Kloning dan transformasi vektor PJ804:77539 dilakukan dengan tujuan perbanyakan vektor pRHA pada sel bakteri E.coli TOP’10. Ekpresi vektorpRHA diharapkan dapat terjadi pada periplasma E.coli dan memberikan ekspresi berupa kemampuan resistensi terhadap Ampicillin. Ekspresi pada periplasma bertujuan untuk meminimalisir kerugian yang timbul pada sistem ekspresi di sitoplasma di antaranya tingkat ekspresi yang rendah, protein terpotong atau resiko kontaminasi. Sekresi protein rekombinan pada periplasma dapat meningkatkan aktivitas biologis serta tingkat kestabilan produk menjadi lebih tinggi. Proses isolasi protein yang diekspresikan pada periplasma  dapat dilakukan dengan perlakuan stress osmotik ringan sehingga menurunkan resiko kontaminasi protein sitoplasma. Ekspresi protein pada periplasma diarahkan oleh peptida sinyal pelB. Peptida sinyal bekerja menarik produk protein ke periplasma dengan cara berfusi dengan ujung N-terminal pada peptida yang terekspresi. Penanda selektif (selectable marker) yang terdapat pada PJ804::77539 merupakan Ampr, suatu penanda yang memampukan bakteri untuk resisten pada keberadaan antibiotik Ampicillin. Transformasi dilakukan sesuai dengan metode heat – shock dan diseleksi pada medium LB agar dan LB cair yang mengandung antibiotik Ampicillin dengan konsentrasi 100 mg/mL. Diperoleh koloni tumbuh pada medium yang mengandung Ampicillin dan dilakukan isolasi plasmid. Visualisasi hasil elektroforesis memperlihatkan adanya pita plasmid yang diisolasi dari E.coli TOP’10 pada gel elektroforesis.Kata kunci : Ampicillin, E.coli, pelB, periplasm dan pRHA

AKTIVITAS SPESIFIK SELULASE Serratia marcescens DENGAN VARIASI KONSENTRASI AMONIUM SULFAT ((NH4)2SO4) DAN pH

Jurnal Akademika Biologi Vol. 6 No. 2 April 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Cellulose is a component found in the cellular structure in almost all plant matter, its existence considered to be the most abundant on earth, and even excreted by some bacteria. Cellulose degradation is performed by cellulase enzymes consisting of three components, namely, endoglucanase, exoglucanase, and β-glucosidase enzyme with glucose as the final product. Cellulase utilization is often used in the textile, food and paper industries, whereas in the field of pharmaceuticals, cellulase enzymes are used to maintain optimal digestive health or produce substances that act as binding tablets such as methylcellulose, ethylcellulose, and hydroxypropylcellulose. The purpose of this study is to determine the concentration of ammonium sulfate and optimum pH for cellulose specific activity of Serratia marcescens. Determination of cellulase activity was done by DNS method, while determination of protein content was done by Lowry method. This research uses Randomized Block Design (RAK) factorial pattern with two factors. The first factor was variation of ammonium sulfate concentration which consisted of (0%, 0,75% and 1%). The second factor is the variation of pH consisting of 6, 7, and 8. Each factor is repeated 3 times. The data obtained were analyzed using Analysis Of Variance (ANOVA). The results showed that the combination of ammonium sulphate concentration variation with pH was not optimum to increase cellulose specific activity of S. marcescens.    Keywords: Cellulase, Ammonium sulfate, pH, Serratia marcescens

PENGARUH CaCl2.2H2O DAN WAKTU INKUBASI TERHADAP PRODUKSI INULINASE OLEH Pichia manshurica DUCC Y-015 DALAM SUBSTRAT TEPUNG UMBI DAHLIA

Jurnal Akademika Biologi Vol. 6 No. 3 Juli 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Fructose production from inulin by inulinase only need one reaction enzimatic step and produce 95% fructose. Inulin obtained from dahlia tuber and inulinase produced by Pichia manshurica DUCC Y-015. Inulinase production (E.C. 3.2.1.7.) can be influenced by metal salt suplementation, such as CaCl2.2H2O. The purpose of this research were to known the influence of CaCl2.2H2O and incubation time to inulinase production by P. manshurica DUCC Y-015 on Dahlia Tuber Substrate. The design that use in this research were Randomized Factorial Block Design ( RAFBD ). Factor I (CO, C­1, C2, C3) as the concentration of CaCl2.2H2O (0 mM, 0.25 mM, 0.50 mM, 1.00 mM) and Factor II ( T12, T18, dan T24 ) as incubation time ( 6, 12, 18 hour), the repetition were 3 times. The result analyze by ANOVA (Analysis of Variance) and continued by LSD test. The result of this research indicate that CaCl2.2H2O and incubation time were not significantly influence to inulinase production. The highest inulinase production by Pichia manshurica DUCC Y-015 indicate by C2T12 treatment which use 0.50 mM CaCl2.2H2O and 12 hour incubation time, the enzyme activity is 0.60 IU/mLKey Words : CaCl2.2H2O, Dahlia tuber,  Incubation time, Inulin, Inulinase, Pichia manshurica DUCC Y-015.

IDENTIFIKASI MOLEKULER TANAMAN PISANG RAJALAWE BERDASARKAN GEN INTERNAL TRANCRIBED SPACER (ITS)

Jurnal Akademika Biologi Vol. 6 No. 1 Januari 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Banana is one type of horticultural commodities in a group of fruits that have a socio-economic value is high enough for the people of Indonesia. Bananas have different varieties, one banana type Rajalawe found in Central Java. Rajalawe molecular identification has not been done before. This study aims to determine the result of the identification of the molecular basis of  Rajalawe based on genes Internal transcribed spacer (ITS), in search of identity and kinship Rajalawe. The study was conducted by isolating DNA using a method Rajalawe Doyle & Doyle, followed by ITS gene amplification and sequencing analysis. The results of gene amplification ITS produce PCR product of 643 bp. The base sequence of the sequencing results are used for the construction of phylogenetic trees. Sequence similarity analysis Rajalawe show 95% homology with Musa balbisiana and alkaline difference of 1%. Phylogenetic tree analysis showed Rajalawe have a close relationship with Musa balbisiana. However, bananas Rajalawe has several different characters with Musa balbisiana with different base sequences by 5% whereas the base sequence homology between the banana Musa balbisiana and Rajalawe with 95%.Keywords: Molecular Identification, Pisang Rajalawe, Universal Primer ITS, Musa balbisiana.

PENAPISAN DAN PEMANFAATAN RHIZOBAKTERI TANAMAN SORGUM (Sorghum bicolor L. Moench) SEBAGAI INOKULAN PEMACU TUMBUH TANAMAN

Jurnal Akademika Biologi Vol. 5 No. 4 Oktober 2016
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Sorghum was a cereal crop that has many benefit such as food, feed, industrial, and bioenergy. Sorghum had a potency to be cultivated, but productivity of sorghum was still low both in quatity and quality. One way to increase production of sorghum  is using rhizobacteria as biofertilizer. The aim of this study is to get rhizobacteria that has the ability to produce IAA, solubility of phospat (P), Nitrogen (N) fixing, and analyze the effect of rhizobacteria inoculants for enhance sorgum plant growth. Isolation of rhizobacteria was done by diluting  rhizobacteria sorghum suspension from 10-1 to 10-5 and it were be platted on SEA medium. Isolates were screened by ability to produce IAA, solubility of P, and N fixing. Producing of IAA test was done by adding Salkowsky reagent on bacterial supernatant and measured absorbance at 530 nm wavelength. Solubility of P test was done by inoculating isolates in Pikovskaya media, while N fixing test was done on N fixing media (NFB). Isolates of rhizobacteria which had a potency to increase growth of plants were made inoculants to be applied in sorghum plants. The result of this study obtain 3 isolates i.e Sr 194.3; Sr 172.1; and Sr 209.1 which were considered effective for increase growth of sorghum. The conclusion  of this study isolates which showed the highest average plant height, root length, and dry weight Sr 194.3 isolate. The statistical analysis among the treatments showed that did not any significant differences on plant height, root length, and dry weight of sorghum age 28 days after farming. Keyword : Increase growth plants, Screening, Shorgum, Rhizophere.

ISOLASI DAN IDENTIFIKASI BAKTERI GENUS Sphingomonas DARI DAUN PADI (Oryza sativa) DI AREA PERSAWAHAN CIBINONG

Jurnal Akademika Biologi Vol. 6 No. 1 Januari 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

The unique ability of  the genus Sphingomonas bacteria as degrade the contaminants refractory contaminants, to serve as the antagonists bacteria to phytopathogenic fungi, and capable to secrete  hidhly useful exopolysaccharide gellan make these bacteria may play an important role in various industrial fields. Exploitation of the metabolic capabilities by genus Sphingomonas bacteria can provide significant commercial advantages for biotechnology.The species of Sphingomonas are often found associated with the rice plant as one of the endophytic bacteria that can be cultured. This study aims to isolate the local bacteria that can produce gellan gum from the leaves of the rice plant (Oryza sativa). The isolation process is done with a spread plate method suspension of rice leaves on Nutrient Dextrose Agar (NDA) media. Single colonies of bacteria that can be isolated then identified by colony PCR method to proceed at sequencing process. Sequencing followed by equalization sequences on the BLAST program shows four isolates of the genus Sphingomonas which isolates XA1, XA2, XA6, XA12 with the results are Sphingomonas sp. Fse41, Sphingomonas sp. Fse41, Sphingomonas sanguinis L4-317 strain and Sphingomonas sp. MLB01Keywords: endophytic bacteria, padi, Sphingomonas

PRODUKSI ENZIM INULINASE Pichia manshurica DUCC-Y015 DENGAN PENAMBAHAN SUBSTRAT TEPUNG BENGKOANG (Paschyrhizus erosus)

Jurnal Akademika Biologi Vol. 6 No. 4 Oktober 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Bengkoang (Pachyrhizus erosus) tubers has a high inulin content. Inulin bengkoang flour can be used as substrate to produce inulinase enzyme. The inulinase enzyme can be produced by Pichia manshurica DUCC-Y015. This research aims to determine the ability of Pichia mansurica DUCC-Y015 in producing inulinase enzyme with the addition of several variations of substrate concentration of bengkoang flour in its production medium. Determination of inulinase activity was done by DNS method. This research used a Completely Randomized Design (RAL) with 4 treatments: B0 (control), B1 (1 g bengkoang flour), B2 (3 g bengkoang flour) and B3 (5 g bengkoang flour). Each treatment was repeated 3 times. The inulinase activity of each treatment was 0.029 IU/mL, 0.033 IU/mL, 0.053 IU/mL and 0.015 IU/mL. The addition of variation substrate concentration bengkoang flour in the production medium did not affect the inulinase activity of Pichia manshurica DUCC-Y015Kata Kunci: Pachyrhizus erosus, inulinase, Pichia manshurica DUCC-Y015.

AKTIVITAS ENZYM SELULASE YANG DIHASILKAN OLEH BAKTERI Serratia marcescens PADA SUBSTRAT JERAMI

Jurnal Akademika Biologi Vol. 7 No. 1 Januari 2018
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Cellulose (EC 3.2.1.4) is enzyme complex consisting of some enzymes which together decomposing cellulose into glucose by hydrolizes the β-1,4 bond in cellulose. The purpose of this study is to determine cellulose activity which produced by Serratia marcescens in different substrate concentration and at the time of incubation T4, T8, T12. This research uses Randomized Block Design (RBD) factorial pattern with two factors. The first factor was variation of straw substrate which had been delignificated (V0, V1, V2, V3). The second factor is the variation of time incubation (T4, T8, T12). Each factor is repeated 3 times. The data obtained were analyzed using Analysis  of  Variance (ANOVA)  (α  =  0.05). The result  shown that  variation concentration  of straw,  and the interaction (combination) between the straw substrate and the incubation time substrate was not significantly different. The result treatment of incubation time was significantly different of the cellulase activity. The result of anova analyzed is obtained that F count(α = 0.05) value from straw substrate, interaction (combination) between the straw substrate and the incubation time substrate, and incubation time was 0.53; 2.18; 8.00. F table(α = 0.05) value of straw substrate, interaction (combination) between the straw substrate and the incubation time substrate, and incubation time was 2.99; 2.20; 3.39. The result of anova, is continued by BNT 5% test. The result of BNT test shown that the highest incubation time of cellulase activity was in incubation time 12 hours with the average value 0.26 U/mL. Key Word : cellulose,  Serratia marcescens,  straw substrate, incubation time