Pengaruh proporsi tepung labu kuning dan tepung terigu terhadap sifat fisikokimia dan organoleptik cake kukus

Labu kuning kaya akan β-karoten, gula, dan serat. Upaya diversifikasi pengolahan labu kuning adalah tepung labu kuning. Tepung labu kuning dapat dimanfaatkan dalam pembuatan cake kukus sebagai bahan pensubstitusi tepung terigu. Penggunaan tepung labu kuning diharapkan dapat meningkatkan kadar β-karoten dalam cake kukus. Penelitian ini bertujuan untuk menentukan proporsi tepung labu kuning dan tepung terigu yang tepat untuk menghasilkan cake kukus dengan sifat fisikokimia dan organoleptik yang dapat diterima oleh konsumen. Rancangan penelitian yang digunakan adalah Rancangan Acak Kelompok dengan satu faktor, yaitu faktor proporsi tepung labu kuning dan tepung terigu dengan lima level, yaitu: 0%:100%; 10%:90%; 20%:80%; 30%:70%; 40%:60%, yang diulang sebanyak lima kali. Parameter pengujian meliputi volume spesifik, firmness, dan organoleptik kesukaan warna, kemudahan digigit, moistness, dan rasa. Data dianalisis dengan ANOVA (Analysis of Variance) pada α = 5% untuk mengetahui ada tidaknya pengaruh proporsi tepung labu kuning dan tepung terigu terhadap sifat fisikokimia dan organoleptik cake kukus. Apabila terdapat pengaruh nyata maka dilanjutkan dengan uji DMRT (Duncan's Multiple Range Test) pada α = 5%. Penentuan perlakuan terbaik dilakukan dengan uji pembobotan. Cake kukus dengan perlakuan terbaik diuji kadar β-karoten. Hasil penelitian menunjukkan bahwa perlakuan proporsi tepung labu kuning : tepung terigu memberikan pengaruh nyata (α = 5%) terhadap volume spesifik, firmness, dan tingkat kesukaan terhadap warna, kemudahan digigit, moistness, dan rasa. Semakin besar proporsi tepung labu kuning, semakin kecil nilai volume spesifik, firmness, dan kesukaan warna cake, namun semakin besar nilai kesukaan kemudahan digigit, moistness, dan rasa cake. Proporsi tepung labu kuning : tepung terigu 20%:80% merupakan perlakuan terbaik dengan kadar β-karoten sebesar 8,569 µg β- karoten/g cake kukus

Summary table.

<p>*- indeterminate</p><p>**—maternal anti-HEV IgG; n.a.-not available; n.d.-not done.</p><p>HEV in minipigs.</p

Group 3.

<p>n.t.- not tested.</p><p>Fatteners (~ 10 months old). Analysis of sera (s).</p

Microorganisms tested negative in three “retired breeders” minipigs, age 24–48 months.

<p>Microorganisms tested negative in three “retired breeders” minipigs, age 24–48 months.</p

Group 1.

<p>Adult minipigs “retired breeders”. Analysis of sera (s) and islets cells (i).</p

HEV genome, primers and probes, recombinant proteins and real-time RT-PCR parameters.

<p>(A) Schematic presentation of the HEV genome with three open reading frames (ORF 1–3). UTR—untranslated region. The numbers of nucleotides are given from the first nucleotide of ORF 2. Cap- cap structure, Poly (A)—poly A sequence. PCR primers (black arrows) and probes (red arrows) used in the real-time RT-PCR methods “A”, “J”, “M1”,and “M2” for the detection of HEV ORF2. Primers for methods M1 and M2 are given in brackets as 1 or 2, respectively. Numbers are given as in A. ^*Method “A” was established by Adlhoch et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139893#pone.0139893.ref041" target="_blank">41</a>]; **Method “J” was established by Jothikumar et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139893#pone.0139893.ref059" target="_blank">59</a>]. (B) Parameters of the used real-time RT-PCR methods. Standard curves and PCR efficiencies are shown. One reference plasmid was used for the real-time RT-PCR method “A” and another, for the other three methods. (C) HEV ORF 2 is coding for the 660 aa long capsid glycoprotein. Putative glycosylation sites are marked in green, a low probability glycosylation site is marked in grey. Positions of the asparagine are numbered. The immunodominant region (IDR) is given in yellow. The signal peptide is shown as a blue arrow. (D) Recombinant proteins (Prospec and GT3) used as antigens in Western blot analysis are shown. GT3 contains the entire immunodominant region (IDR). The Prospec antigen contains glutathion-S-transferase (GST) on the N-terminus. GT3 contains a 6His-tag on the N-terminus. The fragments corresponding to the ORF2 are given in green. The numbers started from the first amino acids of the capsid protein. (E) Comparative analysis of the recombinant antigens by SDS-PAGE is shown on the right. 500 ng of proteins were loaded on the gel, the gel was stained with Coomassie brilliant blue. Lane 1 –Prospec, lane 2 –GT3, M–Size markers.</p

Western blot analysis of sera from Göttingen Minipigs.

<p>(A) Western blot analysis of sera from retired breeders (Group 1). The numbers on strips corresponded to the order of animals in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0139893#pone.0139893.t002" target="_blank">Table 2</a>. “+”–serum from a HEV infected pig diluted 1:300; “+ (bold)” serum from infected pig diluted 1:150. The antigen load was 300 ng/strip. Animal sera were tested twice in dilution 1:150. (B) Analysis of sera from sow-piglet pairs (Group 4). Odd numbers 1, 3, 5, 7, 9, 11 –strips incubated with sera from sows; even numbers 2, 4, 6, 8, 10, 12 –strips incubated with sera from piglets. Strip 4 was incubated with serum of piglet #319428. Strips 7, 8 (underlined) were treated with serum of the sow #314253 and serum of piglet #320282, respectively. “+” -serum from HEV infected pig; “-” serum from non-infected pig. The antigen load was 300 ng/strip. The sera specimens were diluted 1:150.</p

Group 2.

<p>Adult minipigs. Analysis of kidney (k) and liver (l).</p

Analysis of the HEV sequences revealed in two sows.

<p>(A) Neighbour-joining phylogenetic tree of nucleotide sequences corresponding to a 144 nucleotides region in ORF2. Sequences from the infected sows #31453 and #314451 are given in bold. After the animal number the number of identical clones derived from the same amplicon is given in brackets as (+n). Accession numbers from the GenBank sequences are indicated. The origin of the sequences is indicated as: h-human; s-swine; wb-wild boar; rb-rabbit. The genotype and subgenotype are given as a number and a small letter, respectively. Absence of the letter means that the subgenotype is unknown. Brackets mark the HEV genotypes 3 and 4 (indicated as gt3 and gt4), respectively. The reference (Ref.) plasmid was designed using WHO standard of HEV (Acc. #AB630970). Nucleotide substitutions per 100 nucleotides are indicated. (B) Alignment of deduced amino acid sequences (48 residues) of cloned HEV sequences from sow #314451 and sow #314253 using Clustal W software. Amino acids substitutions associated with reduced infectivity of HEV (underlined in the consensus sequence) were not detected in the cloned sequences.</p

Lack of HEV in liver and kidney tissues from adult Göttingen minipigs.

<p>Three different duplex real-time RT-PCR systems “A”, “J”, and “M2” were used to screen for HEV in liver and tissue of the animals from Group 2 using 500 ng of total RNA for the analysis. Porcine cyclophilin A gene was used as a house keeping control and equal load of RNA was confirmed. Serial 10-fold dilutions (10E5-10E2) of the reference plasmid were used as a control for the system “J”. Analyses were performed in duplicates. In addition, minipigs from Group 2 were also found negative when RNA specimens were tested using a conventional semi-nested RT-PCR (data not shown).</p