Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source
The search for eco-friendly alternatives to conventional petroleum-based materials has intensified in an era marked by a growing global awareness of environmental sustainability. This study addresses the critical need for molecular identification and characterization of fungi sourced from a tempeh...
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IIUM Press, International Islamic University Malaysia
2025-05-01
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| Series: | International Islamic University Malaysia Engineering Journal |
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| Online Access: | https://journals.iium.edu.my/ejournal/index.php/iiumej/article/view/3293 |
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| author | Ainil Hawa Jasni Azlin Suhaida Azmi Noor Illi Mohamad Puad Fathilah Ali Yusilawati Ahmad Nor |
| author_facet | Ainil Hawa Jasni Azlin Suhaida Azmi Noor Illi Mohamad Puad Fathilah Ali Yusilawati Ahmad Nor |
| author_sort | Ainil Hawa Jasni |
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The search for eco-friendly alternatives to conventional petroleum-based materials has intensified in an era marked by a growing global awareness of environmental sustainability. This study addresses the critical need for molecular identification and characterization of fungi sourced from a tempeh commercial starter culture for their potential role in fungal-based polymer production. The problem is the limited knowledge and understanding of the genetic composition of these fungi and their suitability for lactic acid (LA) production, which is a crucial component of fungal-based polymer manufacturing. This research examined the tempeh starter culture fungi to identify suitable strains for LA production. The fungi were genotyped by DNA sequencing of the ITS region. The study revealed that the Ragi tempeh commercial starter culture contained only one strain of Rhizopus (R. microsporus), which was verified through ITS rRNA sequencing with 99.8% similarity to the GenBank database, simplifying control over fungal growth and potentially leading to consistent biomaterial yields. The method employed, involving DNA PCR (and sequencing of the ITS region, proved to be accurate, straightforward, and not excessively labor-intensive. The PCR conditions were as follows: initial denaturation at 98°C for 2 min, followed by 25 cycles of denaturation (98°C for 15 seconds), annealing (60°C for 30 seconds), and elongation (72°C for 30 seconds), with a final extension at 72°C for 10 min. Consequently, the consistent presence of only one Rhizopus species in commercial starter cultures of tempeh presents a promising avenue for sustainable biomaterial production, particularly in LA production. The pilot flask setup at 1 × 10? spores/mL was inoculated into 150 mL shake flasks with 1.2 g/mL glucose, incubated at 37°C for 1 to 7 days with 100 rpm shaking, yielding 1.037 g/g after 5 days, demonstrating the feasibility of using this strain for industrial applications.
ABSTRAK: Dalam era yang semakin menekankan kesedaran global terhadap kelestarian alam sekitar, pencarian alternatif mesra alam kepada bahan berasaskan petroleum konvensional semakin giat dijalankan. Kajian ini menangani keperluan kritikal untuk mengenal pasti dan mencirikan kulat secara molekul daripada kultur pemula komersial tempeh bagi potensi penggunaannya dalam penghasilan asid laktik (LA), komponen penting dalam pembuatan polimer berasaskan kulat. Masalah utama yang dibincangkan ialah kekurangan pengetahuan dan pemahaman mengenai komposisi genetik kulat ini serta kesesuaiannya untuk sintesis LA. Kajian ini menumpukan kepada pemeriksaan kulat dari kultur pemula tempeh bagi mengenal pasti strain yang sesuai untuk pengeluaran LA. Penjujukan DNA kawasan internal transcribed spacer (ITS) digunakan untuk mengenal pasti genotip kulat tersebut. Hasil kajian menunjukkan bahawa kultur pemula komersial Ragi tempeh hanya mengandungi satu strain sahaja, iaitu Rhizopus microsporus (disahkan melalui penjujukan ITS rRNA dengan 99.8% kesamaan dengan pangkalan data GenBank). Kehadiran satu spesies ini memudahkan kawalan pertumbuhan kulat dan berpotensi meningkatkan konsistensi hasil pengeluaran. Kaedah yang digunakan melibatkan PCR DNA dan penjujukan kawasan ITS, yang terbukti tepat, mudah, serta tidak terlalu memerlukan tenaga kerja yang banyak. Keadaan PCR adalah seperti berikut: penyahdenaturan awal pada suhu 98°C selama 2 minit, diikuti 25 kitaran yang terdiri daripada penyahdenaturan (98°C, 15 saat), pengannealan (60°C, 30 saat), dan pemanjangan (72°C, 30 saat), dengan pemanjangan akhir pada 72°C selama 10 minit. Kehadiran spesies Rhizopus yang konsisten dalam kultur pemula tempeh komersial membuka peluang yang menjanjikan untuk pengeluaran asid laktik yang mampan. Penggunaan susunan flask perintis pada suhu 30°C menghasilkan 1.037 g/g selepas 5 hari, membuktikan potensi strain ini untuk aplikasi industri.
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| format | Article |
| id | doaj-art-457bdf9363e044f997d66e4a61252f63 |
| institution | OA Journals |
| issn | 1511-788X 2289-7860 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | IIUM Press, International Islamic University Malaysia |
| record_format | Article |
| series | International Islamic University Malaysia Engineering Journal |
| spelling | doaj-art-457bdf9363e044f997d66e4a61252f632025-08-20T02:30:58ZengIIUM Press, International Islamic University MalaysiaInternational Islamic University Malaysia Engineering Journal1511-788X2289-78602025-05-0126210.31436/iiumej.v26i2.3293Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production SourceAinil Hawa Jasni0https://orcid.org/0000-0002-1013-1368Azlin Suhaida Azmi1https://orcid.org/0000-0001-8841-8273Noor Illi Mohamad Puad2https://orcid.org/0000-0001-8127-0085Fathilah Ali3Yusilawati Ahmad Nor4International Islamic University MalaysiaInternational Islamic University MalaysiaInternational Islamic University MalaysiaInternational Islamic University MalaysiaInternational Islamic University Malaysia The search for eco-friendly alternatives to conventional petroleum-based materials has intensified in an era marked by a growing global awareness of environmental sustainability. This study addresses the critical need for molecular identification and characterization of fungi sourced from a tempeh commercial starter culture for their potential role in fungal-based polymer production. The problem is the limited knowledge and understanding of the genetic composition of these fungi and their suitability for lactic acid (LA) production, which is a crucial component of fungal-based polymer manufacturing. This research examined the tempeh starter culture fungi to identify suitable strains for LA production. The fungi were genotyped by DNA sequencing of the ITS region. The study revealed that the Ragi tempeh commercial starter culture contained only one strain of Rhizopus (R. microsporus), which was verified through ITS rRNA sequencing with 99.8% similarity to the GenBank database, simplifying control over fungal growth and potentially leading to consistent biomaterial yields. The method employed, involving DNA PCR (and sequencing of the ITS region, proved to be accurate, straightforward, and not excessively labor-intensive. The PCR conditions were as follows: initial denaturation at 98°C for 2 min, followed by 25 cycles of denaturation (98°C for 15 seconds), annealing (60°C for 30 seconds), and elongation (72°C for 30 seconds), with a final extension at 72°C for 10 min. Consequently, the consistent presence of only one Rhizopus species in commercial starter cultures of tempeh presents a promising avenue for sustainable biomaterial production, particularly in LA production. The pilot flask setup at 1 × 10? spores/mL was inoculated into 150 mL shake flasks with 1.2 g/mL glucose, incubated at 37°C for 1 to 7 days with 100 rpm shaking, yielding 1.037 g/g after 5 days, demonstrating the feasibility of using this strain for industrial applications. ABSTRAK: Dalam era yang semakin menekankan kesedaran global terhadap kelestarian alam sekitar, pencarian alternatif mesra alam kepada bahan berasaskan petroleum konvensional semakin giat dijalankan. Kajian ini menangani keperluan kritikal untuk mengenal pasti dan mencirikan kulat secara molekul daripada kultur pemula komersial tempeh bagi potensi penggunaannya dalam penghasilan asid laktik (LA), komponen penting dalam pembuatan polimer berasaskan kulat. Masalah utama yang dibincangkan ialah kekurangan pengetahuan dan pemahaman mengenai komposisi genetik kulat ini serta kesesuaiannya untuk sintesis LA. Kajian ini menumpukan kepada pemeriksaan kulat dari kultur pemula tempeh bagi mengenal pasti strain yang sesuai untuk pengeluaran LA. Penjujukan DNA kawasan internal transcribed spacer (ITS) digunakan untuk mengenal pasti genotip kulat tersebut. Hasil kajian menunjukkan bahawa kultur pemula komersial Ragi tempeh hanya mengandungi satu strain sahaja, iaitu Rhizopus microsporus (disahkan melalui penjujukan ITS rRNA dengan 99.8% kesamaan dengan pangkalan data GenBank). Kehadiran satu spesies ini memudahkan kawalan pertumbuhan kulat dan berpotensi meningkatkan konsistensi hasil pengeluaran. Kaedah yang digunakan melibatkan PCR DNA dan penjujukan kawasan ITS, yang terbukti tepat, mudah, serta tidak terlalu memerlukan tenaga kerja yang banyak. Keadaan PCR adalah seperti berikut: penyahdenaturan awal pada suhu 98°C selama 2 minit, diikuti 25 kitaran yang terdiri daripada penyahdenaturan (98°C, 15 saat), pengannealan (60°C, 30 saat), dan pemanjangan (72°C, 30 saat), dengan pemanjangan akhir pada 72°C selama 10 minit. Kehadiran spesies Rhizopus yang konsisten dalam kultur pemula tempeh komersial membuka peluang yang menjanjikan untuk pengeluaran asid laktik yang mampan. Penggunaan susunan flask perintis pada suhu 30°C menghasilkan 1.037 g/g selepas 5 hari, membuktikan potensi strain ini untuk aplikasi industri. https://journals.iium.edu.my/ejournal/index.php/iiumej/article/view/3293DNA sequencing of ITS regionDNA PCR sequencingFungal-based polymer productionlactic acid productionmolecular identification of fungi |
| spellingShingle | Ainil Hawa Jasni Azlin Suhaida Azmi Noor Illi Mohamad Puad Fathilah Ali Yusilawati Ahmad Nor Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source International Islamic University Malaysia Engineering Journal DNA sequencing of ITS region DNA PCR sequencing Fungal-based polymer production lactic acid production molecular identification of fungi |
| title | Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source |
| title_full | Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source |
| title_fullStr | Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source |
| title_full_unstemmed | Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source |
| title_short | Identification of the Rhizopus sp. Fungi as an Alternative Lactic Acid Production Source |
| title_sort | identification of the rhizopus sp fungi as an alternative lactic acid production source |
| topic | DNA sequencing of ITS region DNA PCR sequencing Fungal-based polymer production lactic acid production molecular identification of fungi |
| url | https://journals.iium.edu.my/ejournal/index.php/iiumej/article/view/3293 |
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