การคัดแยกและจำแนกยีสต์สะสมไขมันสูงจากดินบริเวณอ่างเก็บน้ำบางพระ จังหวัดชลบุรี
Isolation and Identification of Oleaginous Yeasts from Soils at Bang Phra Reservoir, Chonburi Province
Keywords:
ไขมันจากจุลินทรีย์ , ยีสต์สะสมไขมันสูง , Candida, Cyberlindnera, SchwanniomycesAbstract
ไขมันจากยีสต์สะสมไขมันสูงสามารถนำไปใช้ประโยชน์ได้หลายด้านรวมทั้งการผลิตไบโอดีเซลซึ่งเป็นพลังงานทางเลือกประเภทหนึ่งที่ได้รับความสนใจในปัจจุบัน งานวิจัยนี้จึงมีวัตถุประสงค์เพื่อคัดแยก และจัดจำแนกยีสต์ที่มีศักยภาพในการสะสมไขมันสูงจากดินที่เก็บรวบรวมจากบริเวณอ่างเก็บน้ำบางพระ จังหวัดชลบุรี พบว่าสามารถคัดแยกยีสต์ได้ทั้งหมด 39 ไอโซเลท และเมื่อนำยีสต์ทั้งหมดมาคัดกรองการสะสมไขมันด้วยวิธี Nile red fluorescence assay และตรวจสอบการสะสมไขมันของยีสต์ด้วยวิธีย้อมสี Sudan Black B พบว่ามียีสต์ 8 ไอโซเลท ได้แก่ I2, J1, J2, J5, L1, L4, M2 และ O3 ที่มีศักยภาพในการสะสมไขมันสูง เมื่อนำยีสต์ทั้ง 8 ไอโซเลทมาจัดจำแนกด้วยการวิเคราะห์ลำดับนิวคลีโอไทด์บริเวณ ITS1-5.8S rDNA-ITS2 ร่วมกับการวิเคราะห์สายสัมพันธ์เชิงวิวัฒนาการ พบว่ายีสต์ไอโซเลท I2, J5 และ L1 จัดจำแนกได้เป็น Candida tropicalis ยีสต์ไอโซเลท J1 และ M2 จัดจำแนกได้เป็น Cyberlindnera sp. ยีสต์ไอโซเลท J2 และ O3 จัดจำแนกได้เป็น Schwanniomyces vanrijiae var. vanrijiae ในขณะที่ยีสต์ไอโซเลท L4 จัดจำแนกได้เป็น Schwanniomyces sp. โดยยีสต์ ทั้ง 3 จีนัสจัดอยู่ในไฟลัมเดียวกันคือ Ascomycota และแบ่งออกเป็น 2 แฟมิลี โดย Candida กับ Schwanniomyces จัดอยู่ในแฟมิลีเดียวกัน คือ Debaryomycetaceae และ Cyberlindnera จัดอยู่ในแฟมิลี Phaffomycetaceae งานวิจัยนี้แสดงให้เห็นว่าดินเป็นแหล่งอาศัยที่น่าสนใจของยีสต์ที่มีศักยภาพในการสะสมไขมันสูง อีกทั้งให้ข้อมูลพื้นฐานที่สำคัญสำหรับนำไปใช้ในการศึกษาเพิ่มเติมเพื่อนำยีสต์ไปใช้ประโยชน์ต่อไป Lipid from oleaginous yeasts could be exploited for several applications including the production of biodiesel which is an alternative energy that has been gaining much interest recently. This research aimed to isolate and identify potential oleaginous yeasts from soil samples collected at Bang Phra reservoir in Chonburi province. A total of 39 isolates of yeast were obtained and further screened for intracellular lipid production by Nile red fluorescence assay followed by Sudan Black B staining to observe lipid accumulation in yeast cells. The result showed that there were 8 yeast isolates; I2, J1, J2, J5, L1, L4, M2 and O3 that exhibited high level of intracellular lipid accumulation potential. Identification of yeasts based on nucleotide sequencing of ITS1-5.8S rDNA-ITS2 regions as well as phylogenetic analysis showed that I2, J5 and L1 were identified as Candida tropicalis, J1 and M2 were identified as Cyberlindnera sp., and J2 and O3 were identified as Schwanniomyces vanrijiae var. vanrijiae and L4 was identified as Schwanniomyces sp. These yeast strains belong to the same phylum (Ascomycota) which are divided into 2 families as following; Candida and Schwanniomyces are members of the family Debaryomycetaceae and Cyberlindnera is a member of the family Phaffomycetaceae. This research suggested that soil could be an interesting habitat of potential oleaginous yeasts and this work also provided fundamental and important knowledges for further studies aiming for useful applications.References
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