Isolation and partial characterization of β-galactosidase activity produced by a thermotolerant strain of Kluyveromyces marxianus during growth on lactose-containing media

Brady, Damien; Marchant, Roger; McHale, L.; McHale, Anthony P.

doi:10.1016/0141-0229(94)00115-8

Cited by 28 publications

(31 citation statements)

References 15 publications

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“…While previous studies on the immobilized K. marxianus IMB3 system demonstrate increased ethanol production during growth on media containing other carbohydrate sources, the increases in production usually only become noticeable at later stages in fermentation [13,14]. It should be noted that ethanol production by the free organism in the present study is slightly higher than previously reported for the organism during growth on lactose-containing media [10]. In those previous studies Ca2> was omitted from media and we believe that its inclusion in media used in the present study may have contributed to enhanced -galactosidase activity and thermal stability.…”

Section: Resultscontrasting

confidence: 56%

“…However, in lactose-containing media the yields of ethanol are relatively low and a variety of strategies have been used in order to increase those yields. In our studies it has been shown that supplementation of lactose-containing media with the -galactosidase activity derived from K. marxianus IMB3 results in increasing the yield from 30% to 53% of the maximum theoretical yield [10]. It was subsequently discovered that addition of Mn2> to media, supplemented with the K. marxianus IMB3 -galactosidase, contributed to a further increase in the yield of ethanol [11].…”

Section: Resultsmentioning

confidence: 72%

“…As mentioned above the thermotolerant yeast strain K. marxianus IMB3 is capable of growth and ethanol production at 45°C on lactose-containing media [10]. However, in lactose-containing media the yields of ethanol are relatively low and a variety of strategies have been used in order to increase those yields.…”

Section: Resultsmentioning

confidence: 99%

“…Relatively recently we have described a strain of Kluyveromyces marxianus which is capable of producing ethanol at temperatures ranging from 45-50°C from a variety of carbohydrates including lactose [7,8,9,10]. During growth of that strain on lactose containing media at 45°C, it was found that although the organism produced ethanol, maximum yields were relatively low [10].…”

Section: Introductionmentioning

confidence: 99%

“…During growth of that strain on lactose containing media at 45°C, it was found that although the organism produced ethanol, maximum yields were relatively low [10]. However, when media were supplemented with cell extracts from the organism, which contained -galactosidase, ethanol production increased very significantly [10].…”

Section: Introductionmentioning

confidence: 99%

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Ethanol production at 45°C by alginate-immobilized Kluyveromyces marxianus IMB3 during growth on lactose-containing media

Brady¹,

Nigam²,

Marchant³

et al. 1997

Bioprocess Engineering

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The thermotolerant yeast strain Kluyveromyces marxianus IMB3 was immobilized in calcium alginate and this was used in batch-fed reactor systems to convert lactose (4 g/l) to ethanol. Production of ethanol by the free and immobilized biocatalyst in the presence and absence of Mn2> was com pared. In systems containing the free microorganism in the presence and absence of Mn2>, ethanol increased to a maxi mum of 8 g/l within 40 hours with no significant difference in production by both systems. Ethanol production by the immobilized system in the absence of Mn2> increased to a maximum of 13 g/l within 40 hours and then decreased to 9 g/l within 80 hours. Ethanol production by the immobilized system in the presence of Mn2> increased to 14 g/l within 60 hours and this decreased to 13 g/l at 80 hours. When all systems were re-fed at 80 hours, ethanol production by systems containing the free biocatalyst increased to a maximum of 3 g/l while the immobilized system in the presence of Mn2> increased to a maximum of 12 g/l. Subsequent experiments involving re-feeding the system at shorter time intervals demonstrated that ethanol production by the immobilized system on lactose-containing media at 45°C was far superior to ethanol production by the free biocatalyst.

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Section: Resultscontrasting

confidence: 56%

Section: Resultsmentioning

confidence: 72%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ethanol production at 45°C by alginate-immobilized Kluyveromyces marxianus IMB3 during growth on lactose-containing media

Brady¹,

Nigam²,

Marchant³

et al. 1997

Bioprocess Engineering

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Protoplast transformation of Kluyveromyces marxianus

Lyu

Zhou

et al. 2021

Biotechnology Journal

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The dairy yeast Kluyveromyces marxianus is a promising cell factory for producing bioethanol and heterologous proteins, as well as a robust synthetic biology platform host, due to its safe status and beneficial traits, including fast growth and thermotolerance. However, the lack of high-efficiency transformation methods hampers the fundamental research and industrial application of this yeast. Protoplast transformation is one of the most commonly used fungal transformation methods, but it yet remains unexplored in K. marxianus. Here, we established the protoplast transformation method of K. marxianus for the first time. A series of parameters on the transformation efficiency were optimized: cells were collected in the late-log phase and treated with zymolyase for protoplasting; the transformation was performed at 0 • C with carrier DNA, CaCl 2 , and PEG; after transformation, protoplasts were recovered in a solid regeneration medium containing 3-4% agar and 0.8 M sorbitol. By using the optimized method, plasmids of 10, 24, and 58 kb were successfully transformed into K. marxianus.The highest efficiency reached 1.8 × 10 4 transformants per μg DNA, which is 18-fold higher than the lithium acetate method. This protoplast transformation method will promote the genetic engineering of K. marxianus that requires high-efficiency transformation or the introduction of large DNA fragments.

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Analysis of lactase in lactose intolerance supplements

Stourman

Moore

2018

Biochem Molecular Bio Educ

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Lactase is the enzyme responsible for the digestion of the disaccharide lactose, and deficiency in this enzyme causes the prevalent medical condition lactose intolerance. Management of lactose intolerance can be achieved through the administration of lactase supplements. Lactase is an appropriate platform for advanced enzymatic study because its medical application is a motivator for student learning. The following is an upper‐level biochemistry laboratory sequence that integrates student inquiry and exposure to advanced laboratory techniques. Students investigate three different lactase supplements through experimentation that includes the Bradford assay, SDS‐PAGE, continuous and discontinuous kinetic assays, and zymography. Upon completion of this project, students compile their results and conclusions in a scientifically formatted paper comparing supplement protein content and activity. This safe and inexpensive laboratory project enriches student understanding of key biochemical concepts while mirroring work performed in a realistic research setting. © 2018 International Union of Biochemistry and Molecular Biology, 46(6):652–662, 2018.

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Isolation and partial characterization of β-galactosidase activity produced by a thermotolerant strain of Kluyveromyces marxianus during growth on lactose-containing media

Cited by 28 publications

References 15 publications

Ethanol production at 45°C by alginate-immobilized Kluyveromyces marxianus IMB3 during growth on lactose-containing media

Ethanol production at 45°C by alginate-immobilized Kluyveromyces marxianus IMB3 during growth on lactose-containing media

Protoplast transformation of Kluyveromyces marxianus

Analysis of lactase in lactose intolerance supplements

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