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Abstrak :
Situs aktif CDH pada Phanerochaete chrysosporium terdiri dari dua subsites , subsite C katalitik dan B subsite yang mengikat substrat . Domain flavin pada PcCDH Phanerochaete chrysosporium yang telah bermutasi dalam residu F282 berhasil diekspresikan dalam Escherichia coli . Pergantian Phe282 ke Ala , Asp , dan His mengubah aktivitas enzimatik dan spesifisitas spesifisitas terhadap substrat. PcCDH wild-type berperan dengan efisien dalam mengoksidasi hanya selobiosa dan laktosa, sedangka tiga macam variasi Phe282 mampu menunjukkan aktivitas pada glukosa dan maltosa. Mutan mempertahankan sebagian besar aktivitasnya dengan selobiosa tetapi menunjukkan penurunan terhadap laktosa. Kemampuan untuk mengenali glukosa tersebut memberikan peluang besar untuk aplikasi di multibiosensor.

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The active site of cellobiose dehydrogenase from Phanerochaete chrysosporium is composed of two subsites, a catalytic C subsite and a substrate-binding B subsite. The soluble flavin domain of the Phanerochaete chrysosporium CDH that has mutated in residue F282 was successfully expressed in Escherichia coli. Substitution of Phe282 to Ala, Asp, and His changed its enzymatic activity and altered the enzyme?s substrate specificity. While the wild-type cellobiose dehydrogenase efficiently oxidizes only cellobiose and lactose, the three mutated Phe282 also showed activity to glucose and maltose. Mutant retained most of its activity with cellobiose but greatly decreased with lactose. The ability to recognize glucose provide great opportunities for the application in multibiosensor.

Abstrak :
Kambing Saanen merupakan salah satu kambing perah penghasil susu yang baik. Guna meningkatkan efisiensi reproduksi kambing Saanen, dikembangkan teknologi inseminasi buatan (IB). Salah satu faktor pendukung keberhasilan IB yaitu tersedianya semen beku sesuai kriteria. Tujuan dari penelitian yaitu mengetahui pengencer semen yang optimal untuk semen beku kambing Saanen dengan membandingkan pengencer semen berbasis lesitin (Andromed®) dan liposom (Optixcell®) dengan penambahan maltosa 0,4%. Metode koleksi semen dilakukan dengan menggunakan vagina buatan. Semen kambing Saanen kemudian diperiksa secara makroskopis dan mikroskopis. Teknik penyimpanan semen beku dengan teknik kriopreservasi pada suhu -196ºC selama 10 menit. Evaluasi kualitas semen meliputi motilitas, viabilitas, dan membran plasma utuh (MPU). Hasil yang diperoleh diuji secara statistik dengan uji ANAVA satu faktor kemudian uji Tukey. Hasil penelitian menunjukkan tidak terdapat perbedaan nyata (P>0,05) pada nilai rata-rata persentase motilitas perlakuan Andromed® (50,83±3,76); Andromed® + Maltosa 0,4% (50,83±3,76); Optixcell® (50,83±3,76); Optixcell® + Maltosa 0,4% (48,33±6,06), nilai rata-rata persentase viabilitas Andromed® (54,67±3,50); Andromed® + Maltosa 0,4% (54,50±2,51); Optixcell® (56,50±3,45); Optixcell® + Maltosa 0,4%(52,83±5,78), dan nilai rata-rata persentase MPU spermatozoa Andromed® (56,00±3,80); Andromed® + Maltosa 0,4% (56,50±5,47); Optixcell® (53,83±5,31); Optixcell® + Maltosa 0,4% (53,33±6,06). Hasil penelitian menyimpulkan bahwa pengencer semen liposom menghasilkan kualitas yang sama baik dengan pengencer semen berbasis lesitin dan penambahan maltosa 0,4% pada pengencer semen berbasis liposom dan lesitin tidak berpengaruh terhadap kualitas semen beku kambing Saanen yang dihasilkan ......The Saanen goat is a notable domestic goat that performs nicely in dairy production, such as milk. Efforts to improve the reproduction efficiency of Saanen goats, include the development of artificial insemination (IB) technology. A significant factor that effect AI is the availability of frozen semen according to satisfactory criteria. This research aims to determine the optimal semen extender for Saanen goat frozen semen by comparing lecithin‒based extender (Andromed) and liposomes (Optixcell) when added 0.4% maltose. Semen collection utilises artificial vagina, then is examined macroscopically and microscopically. The semen storage technique used in this study was the cryopreservation technique at -196 for 10 minutes. Criteria of semen quality includes motility, viability, and intact plasma membrane. The result obtained were statistically tested with the ANAVA one way factor test then Tukey test. The result showed that there was no significant difference (P>0.05) in the average value of the motility percentage of treatments Andromed (50.83±3.76); Andromed + Maltosa 0.4% (50.83±3.76); Optixcell (50.83±3.76); Optixcell + Maltosa 0.4% (48.33±6.06), viability Andromed (54.67±3.50); Andromed + Maltosa 0.4% (54.50±2.51); Optixcell (56.50±3.45); Optixcell + Maltosa 0.4% (52.83±5.78), and intact plasma membrane Andromed® (56.00±3.80); Andromed + Maltosa 0.4% (56.50±5.47); Optixcell (53.83±5.31); Optixcell + Maltosa 0.4% (53.33±6.06). The results of the study concluded that the liposomes extender produced the same quality as the lecithin-based extender and the addition of 0.4% maltose in liposomes and lecithin‒based extender had no effect on the quality of the frozen semen Saanen goat.

Abstrak :

Kambing Saanen merupakan kambing perah yang dapat menghasilkan susu mencapai 2.695,3 kg dalam satu masa laktasi. Salah satu upaya untuk meningkatkan efisiensi reproduksi serta meningkatkan kualitas kambing perah yaitu penerapan teknik Inseminasi Buatan (IB). Dalam proses pembekuan semen untuk inseminasi buatan, pengenceran semen dan pemberian krioprotektan berpengaruh terhadap kualitas semen beku yang dihasilkan. Penelitian ini bertujuan untuk mengetahui pengaruh penambahan maltosa dengan berbagai konsentrasi ke dalam pengencer berbasis liposom (Optixcell^®) terhadap kualitas spermatozoa kambing Saanen selama kriopreservasi. Penampungan semen dilakukan dengan metode vagina buatan, lalu dilanjutkan dengan pemeriksaan makroskopis dan mikroskopis. Selanjutnya, semen diproses menjadi semen beku menggunakan nitrogen cair -196°C dan setiap tahap kriopreservasi dilakukan evaluasi secara mikroskopis dengan parameter motilitas, viabilitas, dan membran plasma utuh. Penelitian menggunakan Rancangan Acak Lengkap (RAL) dengan empat perlakuan dan enam ulangan. Empat perlakuan terdiri dari kontrol (OK); Optixcell^®+Maltosa 0,2% (O2); Optixcell^®+Maltosa 0,4% (O4); Optixcell^®+Maltosa 0,6% (O6). Data penelitian diuji menggunakan Analisis Varian (ANAVA) satu faktor. Hasil penelitian menunjukkan tidak adanya perbedaan nyata (P>0,05) pada nilai rata-rata presentase motilitas, viabilitas, dan membran plasma utuh. Penambahan maltosa ke dalam pengencer berbasis liposom (Optixcell^®) tidak memiliki pengaruh dalam mempertahankan kualitas semen beku kambing Saanen.

 

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Saanen goat is a dairy goat that can produce milk up to 2,695.3 kg in one lactation period. One of the way to improve reproductive efficiency and quality of dairy goat by implementing Artificial Insemination (IB) techniques. During the process of freezing semen for artificial insemination, the extender and cryoprotectant agents affect the quality of frozen semen produced. The aim of this study was to examine the effect of maltose addition into liposome based extender (Optixcell^®) on quality of Saanen goat during cryopreservation. Semen was collected using artificial vagina, then immediately evaluated in macroscopic and microscopic. Furthermore, the semen was processed into a frozen semen using liquid nitrogen -196°C and every cryopreservation steps were evaluated microscopically using sperm quality parameters such as motility, viability, and membrane integrity. The experimental design was RAL with four treatments and six repetition. Four treatments consisted of Controls (K); Optixcell^®+Maltose 0,2% (O2); Optixcell^®+Maltose 0,4% (O4); Optixcell^®+Maltose 0,6% (O6). The data were analyzed by one way ANAVA. In conclusion, there was no significant difference (P>0.05) in average percentage of motility, viability, and membrane integrity. Addition of maltose to liposome based extender (Optixcell^®) has no significantly to maintain the quality of frozen semen Saanen goat.

 

Abstrak :
Kambing saanen merupakan jenis kambing penghasil susu yang potensial dikembangkan di Indonesia. Keberhasilan inseminasi buatan (IB) dengan semen beku sangat dipengaruhi oleh kualitas semen beku. Penelitian ini bertujuan untuk mengetahui pengaruh penambahan maltosa pada tiga konsentrasi berbeda (0,2%, 0,4%, dan 0,6%) ke dalam pengencer semen berbasis lesitin (Andromed®) terhadap kualitas spermatozoa kambing saanen selama kriopreservasi. Metode penampungan semen menggunakan vagina buatan, semen kambing saanen diperiksa secara makroskopis dan mikroskopis. Teknik penyimpanan semen beku dengan teknik kriopreservasi pada suhu -196ºC selama 10 menit. Evaluasi kualitas semen meliputi viabilitas dan membran plasma utuh. Hasil yang diperoleh diuji secara statistik dengan uji ANAVA satu faktor. Hasil penelitian menunjukkan tidak terdapat perbedaan nyata (P>0,05) pada nilai rata-rata persentase viabilitas dan membran plasma utuh pascathawing. Nilai rata-rata persentase viabilitas pascathawing pada perlakuan Andromed® + Maltosa 0% (54,67±3,50%); Andromed® + Maltosa 0,2% (53,57±3,88%); Andromed® + Maltosa 0,4% (54,50±2,51%); Andromed® + Maltosa 0,6% (51,67±3,39%). Nilai rata-rata persentase membran plasma utuh pascathawing pada perlakuan Andromed® + Maltosa 0% (56,00±3,80%); Andromed® + Maltosa 0,2% (55,50±4,23%); Andromed® + Maltosa 0,4% (56,50±5,47%); Andromed® + Maltosa 0,6% (54,83±3,35%). Hasil Penelitian menyimpulkan bahwa penambahan maltosa dalam pengencer semen berbasis lesitin (Andromed®) tidak memiliki pengaruh dalam mempertahankan kualitas spermatozoa kambing saanen selama kriopreservasi. ......Saanen goat is a type of goat that produces milk that potential to be developed in Indonesia. The success of artificial insemination (IB) with frozen semen is strongly influenced by the quality of frozen semen. This study aims to determine the effect of adding maltose at three different concentrations (0.2%, 0.4%, and 0.6%) to lecithin-based cement diluent (Andromed®) on the quality of saanen goat spermatozoa during cryopreservation. The semen collection method used an artificial vagina, and the semen of the saanen goat was examined macroscopically and microscopically. The technique for frozen semen storage was cryopreservation at -196ºC for 10 minutes. Evaluation of semen quality included viability and intact plasma membrane. The results obtained were tested statistically by one factor ANOVA test. The results showed that there was no significant difference (P> 0.05) in the mean value of the percentage of viability and intact plasma membrane post-thawing. The mean value of the percentage of post-thaw viability in the Andromed® + Maltose 0% (54.67 ± 3.50%); Andromed® + Maltose 0.2% (53.57 ± 3.88%); Andromed® + Maltose 0.4% (54.50 ± 2.51%); Andromed® + Maltose 0.6% (51.67 ± 3.39%). The mean value of post-thawing intact plasma membrane percentage in Andromed® + Maltose 0% (56.00 ± 3.80%); Andromed® + Maltose 0.2% (55.50 ± 4.23%); Andromed® + Maltose 0.4% (56.50 ± 5.47%); Andromed® + Maltose 0.6% (54.83 ± 3.35%). The results of the study conclude that the addition of maltose in lecithin-based cement diluent (Andromed®) did not affect in maintaining the spermatozoa quality of the saanen goat during cryopreservation.

Abstrak :
Ester asam lemak sakarida dapat diperoleh melalui reaksi esterifikasi antara maltosa dengan asam lemak minyak sawit menggunakan pelarut organik. Reaksi esterifikasi dilakukan secara enzimatik menggunakan katalis lipase Candida rugosa E.C.3.1.1.3 terimobilisasi pada nanopartikel Fe3O4-Kitosan, dengan pelarut isoamil alkohol dan iso butanol. Imobilisasi lipase Candida rugosa pada nanopartikel Fe3O4-Kitosan menggunakan metode ikat silang dengan glutaraldehida sebagai agen pengikat silang. Imobilisasi dilakukan menggunakan larutan lipase dengan konsentrasi awal enzim yaitu 200 dan 350 ppm. Variasi yang dilakukan menggunakan perbandingan rasio mol substrat gula: asam lemak, yaitu 1:30 ; 1:60 ; dan 1:90. Hasil yang lebih baik diperoleh menggunakan konsentrasi awal lipase 200 ppm, dengan nilai persen loading imobilisasi lipase yang diperoleh adalah 31,28 % dan aktivitas hidrolisis lipase terimobilisasi sebesar 2,88 U/mL dan aktivitas spesifiknya sebesar 1,23 U/mg. Hasil optimasi perbandingan rasio molar gula dan asam lemak menunjukkan kondisi optimum diperoleh pada rasio 1 : 30 menggunakan pelarut iso butanol, dengan % konversi 5,0 %. Hasil uji emulsifier menunjukkan bahwa produk esterifikasi dapat berfungsi sebagai emulsifier.

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Sugar esters can be produced with esterification between maltose and palm oil fatty acid in organic solvents. Esterification reaction was carried out enzymatically using immobilized Candida rugosa lipase EC.3.1.1.3 on Fe3O4--chitosan nanoparticles, with isoamyl alcohol and isobutanol as the solvents. Candida rugosa lipase was immobilized on Fe3O4-chitosan nanoparticles using cross-linking method with glutaraldehyde as cross linker. Immobilization was carried out with two initial enzyme concentrations of 200 and 350 ppm. The variations of molar substrate ratio of sugar : fatty acid used in this study were 1:30, 1:60, and 1:90. The optimal results was obtained using initial concentration of 200 ppm lipase. The loading percentage of immobilized lipase was 31,28 % with hydrolytic activity of immobilized lipase was 2,88 U/mL and the specific activity was 1,23 U/mg. The highest % conversion in esterification obtained in this study was 5.0 %, using molar ratio 1:30 in iso bthanol as solvent. The ester product obtained was then examined by simple emulsion test and can be used as an emulsifying agent.