สปีชีส์ยีสต์ที่พบในลูกแป้งสาโทจากจังหวัดสุรินทร์: ความสามารถในการย่อยแป้งและการผลิตแอลกอฮอล์
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Abstract
Sasithorn Lorroengsil, Thanongsak Praseartsin, Nara Rodboonrit, Nareerat Moonjai and Jidapa Sangswan
รับบทความ: 29 เมษายน 2565; แก้ไขบทความ: 18 กันยายน 2565; ยอมรับตีพิมพ์: 23 กันยายน 2565; ตีพิมพ์ออนไลน์: 22 ธันวาคม
บทคัดย่อ
งานวิจัยนี้มีวัตถุประสงค์เพื่อคัดแยกเชื้อยีสต์จากลูกแป้งสาโทใน 6 อำเภอของจังหวัดสุรินทร์ ได้แก่ เมืองสุรินทร์ ปราสาท สำโรงทาบ ศรีขรภูมิ ท่าตูมและรัตนบุรี โดยนำตัวอย่างลูกแป้งที่ใช้ในการแยกยีสต์ จำนวน 8 ตัวอย่าง มาเพาะเชื้อในอาหารเหลว YM ที่เติม chloramphenicol ในอัตราส่วน 20 ppm และ sodium propionate 0.05% (น้ำหนัก/ปริมาตร) บ่มที่อุณหภูมิ 35 องศาเซลเซียส เป็นเวลา 24 ชั่วโมง จากนั้นนำมาขีดลากแบบไขว้ (cross streak) บนอาหารแข็ง YM บ่มที่อุณหภูมิ 35 องศาเซลเซียส เป็นเวลา 24–48 ชั่วโมง สามารถแยกยีสต์ได้ทั้งหมด 24 ไอโซเลต เมื่อจัดจำแนกโดยการเทียบลำดับเบสของบริเวณ D1/D2 ของ 26S rDNA ยีสต์จำนวน 20 ไอโซเลต มีผลการระบุชนิดของยีสต์เป็นยีสต์ 4 สปีชีส์ คือ Saccharomycopsis fibuligera (10 ไอโซเลต) Issatchenkia orientalis (5 ไอโซเลต) Pichia kudriavzevii (4 ไอโซเลต) และ Meyerozyma caribbica (1 ไอโซเลต) ซึ่งยีสต์ Sm. fibuligera เป็นสปีชีส์ที่พบบ่อยที่สุด โดยพบในลูกแป้งจำนวน 4 ตัวอย่าง จากทั้งหมด 8 ตัวอย่าง ลูกแป้งส่วนใหญ่พบยีสต์เพียงสปีชีส์เดียวยกเว้น 3 ตัวอย่างที่พบยีสต์ 2 สปีชีส์ โดยลูกแป้งจากอำเภอรัตนบุรีพบยีสต์ Sm. fibuligera ร่วมกับ P. kudriavzeii ลูกแป้งจากอำเภอศรีขรภูมิ (แสงจันทร์) พบยีสต์ Sm. fibuligera ร่วมกับ M. caribbica และลูกแป้งจากอำเภอปราสาท พบยีสต์ I. orientalis ร่วมกับ P. kudriavzevii เมื่อนำยีสต์ทั้ง 24 ไอโซเลตมาทดสอบความสามารถในการย่อยแป้ง พบว่ามียีสต์จำนวน 10 ไอโซเลตที่มีการเจริญและสร้างวงใสบนอาหาร starch agar ได้แก่ ไอโซเลต RTN1–1, RTN1–2, RTN1–3, SRT1–1, SRT1–2, SRT1–3, SRT1–4, SJ1–3, SJ1–4 และ TT1–2 ยีสต์ทั้ง-หมดมีขนาดวงใสใกล้เคียงกันยกเว้น RTN1-1 และ RTN1-3 ที่มีขนาดวงใสเล็กกว่าอย่างมีนัยสำคัญ อย่างไรก็ตามยีสต์จำนวน 10 ไอโซเลต ถูกระบุสปีชีส์เป็นยีสต์ Sm. fibuligera ส่วนความสามารถในการผลิตแอลกอฮอล์ในอาหารเลี้ยงเชื้อ fermentation broth ที่มีน้ำตาลซูโครส 10% (น้ำหนัก/ปริมาตร) พบว่ายีสต์ที่มีประสิทธิภาพในการผลิตแอลกอฮอล์สูงสุด คือ I. orientalis MSR2–2, MSR2–3 และ PS1–5 ซึ่งสามารถผลิตแอลกอฮอล์ได้ 5% (ปริมาตร/ปริมาตร) คิดเป็น 0.39 กรัมเอทานอล/กรัมซูโครส
คำสำคัญ: ยีสต์ ลูกแป้ง สุรินทร์ สาโท
Abstract
The objective of this research was to isolate yeast from the starter (loog–pang) of rice wine (sato) in 6 districts of Surin province including Mueang–Surin, Prasat, Samrong Thap, Sikhoraphum, Thatum and Rattanaburi. Eight samples of loog–pang were used to isolate yeast by adding to YM broth supplemented with 20 ppm of chloramphenicol and 0.05%(w/v) sodium propionate. The flasks were incubated at 35°C for 24 hours. Then yeasts in cultured medium were isolated on YM agar using the cross–streaking method after incubation at 35°C for 24–48 hours. Twenty–four of yeast isolates were obtained in present study. The yeast identification procedure based on the sequencing of amplified D1/D2 region of the yeast 26S ribosomal DNA, twenty of yeast isolates were identified to 4 species including Saccharomycopsis fibuligera (10 isolates), Issatchenkia orientalis (5 isolates), Pichia kudriavzevii (4 isolates) and Meyerozyma caribbica (1 isolate). Yeast Sm. fibuligera was found in 4 samples from total 8 samples which was the most frequently found species in loog-pang. Only one yeast species was found mostly in loog–pang samples, except for 3 samples which two yeast species were found. Yeasts Sm. fibuligera and P. kudriavzeii were found in loog–pang samples from Rattanaburi district, Sm. fibuligera and M. caribbica were found in loog–pang samples from Sikhoraphum (Sang–Jan) district and I. orientalis and P. kudriavzevii were found in loog–pang samples from Prasat district. Among 24 isolates which were tested for starch hydrolysis, it was found that 10 isolates (RTN1–1, RTN1–2, RTN1–3, SRT1–1, SRT1–2, SRT1–3, SRT1–4, SJ1–3, SJ1–4 and TT1–2) could grow and produce clear zone on starch agar. The clear zone diameters of all isolates were similar, except for isolate RTN1–1 and RTN1–3 which were significantly smaller than others. However, all 10 isolates were identified species as Sm. fibuligera. Alcohol production was examined in fermentation broth containing 10%(w/v) sucrose. The highest alcohol contents of 5.0%(v/v) were obtained from I. orientalis isolates MSR2–2, MSR2–3 and PS1–5 with ethanol yield of 0.39 gethanol/gsucrose.
Keywords: Yeast, Loog–pang, Surin, Sato
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