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Article

Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica

by
David Torres-Añorve
and
Georgina Sandoval
*
LIBBA Laboratory, Industrial Biotechnology Unit, Center for Research and Assistance in Technology and Design of the State of Jalisco, A.C. (CIATEJ), Guadalajara 44270, Jalisco, Mexico
*
Author to whom correspondence should be addressed.
Appl. Microbiol. 2025, 5(3), 77; https://doi.org/10.3390/applmicrobiol5030077 (registering DOI)
Submission received: 10 July 2025 / Revised: 28 July 2025 / Accepted: 30 July 2025 / Published: 1 August 2025
(This article belongs to the Special Issue Exclusive Papers Collection of Editorial Board Members and Invited Scholars in Applied Microbiology (2025))
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Abstract

Sustainability represents a significant global challenge, requiring a balance between environmental impact and the use of natural resources. White biotechnology, which uses microorganisms and enzymes for environmentally friendly products and processes, offers promising solutions to support a growing population. Within this context, the yeast Yarrowia lipolytica stands out, so we investigated the generation of biomass from two wild strains (ATCC 9773 and NRRL Y-50997) using different carbon sources. Additionally, protein content and amino acid profiles were assessed via standardized analytical methods to evaluate their potential as nutritional yeasts. Both strains demonstrated potential as nutritional yeasts, with biomass productivities of up to 35.5 g/L and 42 g/L, respectively. The protein content was high, with 58.8% for ATCC 9773 and 58.2% for NRRL Y-50997. Furthermore, the strains presented essential amino acid contents of 62.6% and 41.5%, with lysine being the most abundant amino acid. These findings underscore the versatility and productivity of Y. lipolytica, highlighting its potential for sustainable biotechnological applications such as single-cell protein production.

Graphical Abstract

1. Introduction

One of the current challenges facing humankind is safeguarding its existence through sustainable development. Biotechnology harnesses living organisms to create sustainable products and processes, supporting the development of biobased industries in emerging regions while reducing potential environmental impacts [1]. In 2000, at the meeting of the “Codex Alimentarius Ad Hoc Intergovernmental Task Force on Foods Derived from Biotechnology”, the Food and Agriculture Organization of the United Nations (FAO) recognized that “When appropriately integrated with other technologies for the production of food, agricultural products and services, biotechnology can be of significant assistance in meeting the needs of an expanding and increasingly urbanized population” [2].
In this context, white biotechnology leverages microorganisms and their enzymes to develop environmentally friendly processes, often utilizing agricultural and industrial byproducts for the production of compounds of industrial and nutritional value. For example, in the industrial field, the aim is to reduce the use of fossil fuels by implementing biofuels, to replace conventional energy with bioenergy, and to find ways to accelerate processes in a greener way by using microorganisms that can do this [3]. The environmental application of yeasts also has significant potential, such as bioremediation, where they could contribute to the fight against environmental pollution, as well as in the field of biocontrol to combat plant infections [4]. Yeast cells have the cumulative advantages of high growth capacity, in addition to the extensive knowledge of the genetics of many model species [4].
“Non-Saccharomyces yeasts” or “nonconventional yeasts” (NCY) possess distinctive abilities to produce valuable secondary metabolites. Among the NCY, important genera are Kluyveromyces spp., Pichia spp., and Yarrowia spp. [5]. In particular, Y. lipolytica has notable characteristics, such as “generally recognized as safe” (GRAS) status, versatile metabolism to utilize substrates, flexibility, and metabolic diversity [6]. In addition, it is remarkable for its ability to tolerate extreme environments, such as heavy metal contamination; hypersalinity, acidity, and alkalinity; high biosynthesis and the secretion capabilities of heterologous proteins [7]. Another important feature is the possibility of obtaining high-cell-density cultures, which is of high interest to industries. High-cell-density cultures are a preferable approach to optimize the volumetric productivity of recombinant proteins, biomass, and other components [8]. This parameter is a key factor in the profitability of bioprocesses.
These advantages qualify Y. lipolytica as an ecologically important and valuable candidate for making industrial processes sustainable. Both academia and industry are showing increasing interest in this yeast as a production organism, and this interest is partly driven by metabolic engineering, which has allowed the production of more than 100 different products [9]. Two of the main metabolites of Y. lipolytica, which are excreted extracellularly, are citric acid, which also has a GRAS designation when produced with this yeast, as well as flavorings and sweeteners [10]. In addition, it has lipase 2 (LIP2), which is an enzyme with diverse applications in various fields, including the production of omega-3 concentrates [11], structured dietary lipids [12], and vitamin C derivatives [13] in the food industry; ecological defatting [14], biodiesel synthesis [15], and water bioremediation [16] in the chemical sector; the development of enantiopure drugs [17], treatments for pancreatic insufficiency [18], and bioabsorbable polymers [19] in the pharmaceutical field. Y. lipolytica biomass also has a multitude of applications, as it is rich in proteins and oils. Its biomass has been explored for the production of fragrances [5], supplements [20], carotenoids [21], and antioxidants [22] in the food industry; for bioremediation of uranium [23] and hydrophobic contaminants such as petroleum [24], for water treatment [25], and for the generation of oleochemicals and biofuels [26] in the chemical sector. Additionally, it has the potential to serve as a decontaminating agent when unconventional substrates such as alkanes are consumed.
Furthermore, Y. lipolytica has the capacity to synthesize a range of valuable industrial products from diverse carbon sources, such as glucose and glycerol. For example, certain wild-type strains are efficient at producing single-cell oils, citric acid, acetic acid, kynurenic acid, and polyols from crude glycerol [27,28]. On the other hand, biomass derived from Y. lipolytica cultivated in a medium containing glucose as a carbon source has been utilized as a feed additive for Nile tilapia, resulting in increased nutrient availability and protein and lipid composition in the meat [29], as well as a small effect on the gut microbiome and mucosal immunity in rainbow trout [30].
The ATCC 9773 strain has been used as a reference for several methods [31,32]. Conversely, NRRL Y-50997 can be employed for oil generation from xylose, with the potential to produce biodiesel [33]. However, its utilization for biomass generation has not been extensively investigated. Owing to the various applications that can be applied to Y. lipolytica biomass, the aim of this study was to characterize the biomass production kinetics of a proprietary strain (NRRL Y-50997) in comparison with those of the reference strain Y. lipolytica ATCC 9773 (ATCC 9773) in four culture media. Additionally, the nutritional profile of the biomass was evaluated, as it is relevant for applications such as food additives.

2. Materials and Methods

2.1. Microorganism

The strains of Y. lipolytica used were ATCC 9773 and a strain isolated by the LIBBA Laboratory of the Industrial Biotechnology Unit at CIATEJ named NRRL Y-50997. Both strains were wild type. The strain ATCC 9773 was obtained from the American Type Culture Collection [34]. The yeast NRRL Y-50997 was deposited in the Agricultural Research Service Culture Collection (NRRL) under the Budapest Treaty with the designation NRRL Y-50997 (https://nrrl.ncaur.usda.gov, accessed on 1 June 2025) [33]. The cryogenic strains were stored in 50% (w/w) glycerol at −80 °C.

2.2. Culture Media

The following culture media were prepared from different carbon sources and concentrations: YPD (yeast 10 g/L, peptone 20 g/L, dextrose 20 g/L), YPD6 (yeast 10 g/L, peptone 20 g/L, dextrose 60 g/L), YPG (yeast 10 g/L, peptone 20 g/L, glycerol 20 g/L), and YPG6 (yeast 10 g/L, peptone 20 g/L, glycerol 60 g/L) media. All chemicals utilized were analytical grade. The pH of both culture media was adjusted to 5.0, and the media was sterilized by autoclaving.

2.3. Preparation of Inoculum and Cultivation

For each strain, a preculture was prepared with a cell inoculum in a flask containing YPD medium, and it was cultured at 30 °C with 180 rpm agitation overnight. The strains were subsequently inoculated into 50 mL of YPD medium from each preinoculum and cultured at 30 °C at 180 rpm in a 250 mL flask. Samples were collected from the inoculated media at 0, 6, 12, 24, 48, and 72 h. All microbiological experiments were performed in triplicate.

2.4. Analytical Methods

The growth of the cells was quantified by measuring the dry cell weight (DCW) gravimetrically; for this purpose, the biomass was subjected to a washing process with distilled water, followed by drying for 24 h at 70 °C until a constant weight was achieved, and the resulting dry mass was expressed in grams per liter. The calculations for determining the specific growth rate and biomass-to-substrate yield were conducted in accordance with previously reported methods [35]. Protein and amino acid contents were determined for each strain, which was subsequently grown under standard conditions in YPD media for 24 h. Protein determination followed the NMX-F-608-NORMEX-2011 standard [36]. Amino acids were analyzed via an internal chromatographic method based on the AOAC 982.30 procedure [37]. The total carbohydrate content was estimated via the proximate by difference method [38]; as the final dried biomass contained no residual alcohol, the alcohol term was omitted from this calculation. The fat content was determined in accordance with the 1994 Normative Appendix C, Numeral 1, of the Mexican Official Norm NOM-086-SSA1-1994 [39], whereas the percentage of ash was obtained in accordance with the 2020 Mexican Official Norm NMX-F-607-NORMEX-2020 [40].

3. Results

3.1. Biomass Production

The strains ATCC 9773 and NRRL Y-50997 were cultivated in YPD, YPD6, YPG, and YPG6 media. As illustrated in Figure 1, an analysis of the growth kinetics of strain ATCC 9773 was conducted. Biomass was quantified as dry cell weight in grams per liter over time, with measurements taken at 0, 6, 12, 12, 24, 48, and 72 h. The highest biomass levels were observed at 48 h of incubation in all culture media. Specifically, the highest values of DCW were observed at 48 h, with the following values: YPD = 12.1 g/L, YPD6 = 35.5 g/L, YPG = 20.6 g/L, and YPG6 = 34.2 g/L. These findings suggest that strain ATCC 9773 preferentially utilizes glycerol rather than glucose as a carbon source, as evidenced by the high biomass values observed in glycerol-containing media (YPG and YPG6). Notably, in the YPD medium, a biomass level of 12.1 g/L was observed as early as 24 h, suggesting rapid adaptation and growth of the yeast in the presence of glucose as a carbon source.
In addition, the comparison of the different culture media revealed that the YPD6 medium, which lacks glycerol, presented the highest biomass production at 48 h. This finding indicates that strain ATCC 9773 has a remarkable ability to utilize high concentrations of glucose as a carbon source and to adapt efficiently to high sugar contents in the medium.
The results obtained for strain NRRL Y-50997 in the different culture media exhibited a pattern analogous to that observed with strain ATCC 9773, although with some notable distinctions. Figure 2 depicts the latency, exponential, and stationary phases throughout the incubation period, spanning from 0 to 72 h, across all the evaluated media. The DCW values of YPD, YPG, YPD6, and YPG6 at 48 h were 14 g/L, 27 g/L, 34.5 g/L, and 42.5 g/L, respectively. Furthermore, at 24 h, the biomass generated was 16.5 g/L in the YPD medium and 33.3 g/L in the YPG medium, which is approximately double that of ATCC 9773. These results indicate that NRRL Y-50997 has a predilection for glycerol over glucose as a carbon source, as evidenced by the higher biomass levels observed in the glycerol-containing media (YPG and YPG6). The medium that presented the highest biomass production at 48 h was YPG6, which contained a glycerol concentration of 42 g/L. The difference in DCW between YPD and YPG6 media in the NRRL Y-50997 strain was 28 g/L. This result is consistent with the predilection of the yeast NRRL Y-50997 for glycerol as a carbon source, and since alternative carbon sources have not shown much promise, glycerol is an excellent substrate for the high biomass production of Y. lipolytica.
The maximum biomass, biomass yield to substrate, and specific growth rate are presented in Table 1. A comparison of the biomass obtained by the yeast strains ATCC 9773 and NRRL Y-50997 revealed that NRRL Y-50997 tends to produce relatively high biomass levels. For both YPG and YPG6, NRRL Y-50997 reached significantly higher biomass yields, resulting in biomass productivities above those of ATCC. Moreover, the NRRL Y-50997 strain was superior to the ATCC 9773 strain in terms of biomass-to-substrate yield and specific growth rate on YPG media (YX/S = 1.67 g biomass/g glycerol and μ = 0.15 h−1), whereas the difference in μ was not significant at the elevated glycerol concentration of YPG6, suggesting that excessive substrate partially offsets the kinetic advantage. For both strains, YX/S and μ are greater when glycerol is used instead of glucose.

3.2. Comparison of Biomass Composition

The protein content and composition of the biomass were also investigated. The protein content of Y. lipolytica biomass may vary depending on the carbon source and culture conditions. As shown in Table 2, both strains presented similar protein percentages, with ATCC 9773 (58.8%) and NRRL Y-50997 (58.2%) resulting in a higher protein content than that reported in previous publications for other strains. However, their amino acid compositions differed: strain ATCC 9773 presented an essential amino acid content of 62.6%, whereas strain NRRL Y-50997 presented a content of 41.5%. Although the NRRL Y-50997 strain had a lower percentage of essential amino acids, this was compensated for by the greater biomass yield of this strain. With respect to fat content, strain ATCC 9773 presented a fat content of 3.04%, whereas strain NRRL Y-50997 presented a fat content of 6.35%. Furthermore, strain ATCC 9773 contained 5.80% ash and 32.32% total carbohydrates, whereas NRRL Y-50997 exhibited 6.88% ash and 28.57% total carbohydrates.

3.3. Comparison of Amino Acid Composition

The amino acid compositions of the Y. lipolytica strains ATCC 9773 and NRRL Y-50997 revealed notable differences, particularly in the levels of essential amino acids (Table 3). ATCC 9773 presented the highest lysine (125.1 mg/g protein), leucine (97.8 mg/g), and phenylalanine (72.7 mg/g) contents, whereas NRRL Y-50997 presented elevated levels of arginine (40.5 mg/g) and aromatic amino acids (AAAs: 119.7 mg/g). Both strains presented low cysteine levels (<1.2 mg/g), which were significantly lower than those reported for other protein sources. Compared with the FAO adult requirements, ATCC 9773 exceeded the recommended values for all listed essential amino acids, whereas NRRL Y-50997 met or surpassed most of them, except for isoleucine (22.5 mg/g vs. 30 mg/g FAO) and sulphur amino acids (15.8 mg/g vs. 22 mg/g FAO). Overall, both strains presented amino acid profiles superior to or comparable to those of traditional protein sources such as S. cerevisiae, C. utilis, eggs, and cow milk, highlighting their potential as valuable sources of nutritional protein.
However, the use of Y. lipolytica biomass as an animal feed and dietary supplement has emerged as a promising practice in the agri-food industry (Figure 3). This can be attributed to the fact that the nutritional composition of Y. lipolytica biomass is capable of providing a greater range and diversity of nutritional benefits to the animals that consume it. For this reason, the NRRL Y-50997 strain may be a suitable choice for those seeking to provide their animals with a more diverse range of nutrients.

4. Discussion

The results presented here demonstrate that strains ATCC 9773 and NRRL Y-50997 exhibit distinct metabolic versatility. Importantly, the outcomes observed necessitate further investigation to fully comprehend the potential of each strain. Nevertheless, Y. lipolytica has the potential to contribute to more sustainable and environmentally friendly processes. In particular, the NRRL Y-50997 strain displays promising characteristics for research into environmentally friendly processes and possible use as a food ingredient. During the cultivation of the strains on media with different carbon sources, NRRL Y-50997 was observed to exhibit a predilection for glycerol over glucose, as evidenced by the higher biomass levels observed in the glycerol-containing media. However, an increase in glycerol concentration may result in osmotic stress, which could impede cell growth. For example, previous studies have indicated that following adaptive evolution in a laboratory setting, Y. lipolytica was cultivated in 252.3 g/L (20% v/v) glycerol medium, resulting in the generation of 14.9 g/L dry biomass [52]. Despite greater tolerance to high concentrations of glycerol, the biomass obtained was not greater than that generated by strain ATCC 9773 in YPD6 and YPG6. However, the use of other substrates, such as oat bran, rye bran, and rye straw hydrolysates, has been studied for the growth of strain A-101, where the highest biomass was 9.35 g/L with the use of oat bran [53], without reaching as much biomass as glycerol consumption.
Pure glycerol was used as the carbon source to evaluate the metabolic potential and lipase expression of Y. lipolytica strains. However, it is important to highlight that crude glycerol, a major byproduct of biodiesel production, can also serve as a viable and low-cost substrate for microbial cultivation. The use of crude glycerol not only reduces production costs but also contributes to the valorization of industrial residues within a circular economy framework. Previous studies have demonstrated that Y. lipolytica is capable of efficiently utilizing crude glycerol as the sole carbon source, achieving substantial biomass and lipid accumulation while maintaining a positive energy balance in the overall process [35]. Therefore, the integration of crude glycerol into biotechnological applications with Y. lipolytica represents a sustainable strategy to couple enzyme production with waste management and biofuel industry byproducts [41].
In addition, a previous study demonstrated that the utilization of distinct Y. lipolytica strains isolated from soil yielded comparable biomass levels when cultivated on glycerol-containing media, and when crude or technical glycerol was employed, disparate biomass levels were observed; with crude glycerol resulting in a relatively high biomass yield [54]. It can therefore be inferred that the genetic background of each strain could have a significant effect on growth kinetics and biomass production.
Compared with ATCC 9773, the NRRL Y-50997 strain demonstrated enhanced performance, as reflected by a greater biomass-to-substrate conversion efficiency and a greater specific growth rate when cultivated in the YPG medium. This is consistent with previous reports, which reported a growth rate of 0.3 h−1 for glycerol and 0.24 h−1 for glucose [55]. This characteristic renders NRRL Y-50997 a superior option for the utilization of glycerol, which is a byproduct of numerous industries [56].
The protein content and profile of the Y. lipolytica biomass were also analyzed, revealing that both parameters are influenced by the type of carbon source and the specific cultivation conditions applied. In addition, the higher the nitrogen concentration in the medium is, the greater the amount of protein obtained [57], whereas limiting the amount of nitrogen favors the accumulation of lipids. Notably, both strains presented comparable protein levels, with ATCC 9773 and NRRL Y-50997 yielding protein contents of 58.8% and 58.2%, respectively, values that surpassed those previously reported for other Y. lipolytica strains. For example, the protein content of strain A-101 was significantly lower, at only 19.4%, when it was grown in crude glycerol, and when the same strain was grown on a lipid- and selenium-enriched medium, the resulting biomass contained a significantly greater protein content of 56.4% [28]. Another example is the strain named S11, which produces a small amount of biomass (3.97 g/L), and the protein concentration in the biomass of this strain reached 50.1% when pure glycerol was used [58]. Even when the Y. lipolytica strain CBS 7504 was grown in YPD media to evaluate its use as a dietary supplement for Oncorhynchus mykiss (known as rainbow trout), only a biomass with a nonnegligible protein content of 42.2% was obtained [30]. Peterson et al. reported that co-feeding ethanol with solid cocoa fatty-acid distillates supported cell densities above 40 g/L and a protein fraction just over 45% of dry weight [59]. In contrast, Yang et al. achieved only 24% protein in a one-stage food-waste process but raised this to 38.8% by introducing a two-stage scheme, in which volatile fatty acids generated anaerobically were subsequently converted by Y. lipolytica [60]. Against this backdrop, the 58% protein achieved here for both ATCC 9773 and NRRL Y-50997 on glucose-rich media demonstrated markedly superior protein density, highlighting the nutritional competitiveness of these wild strains despite their reliance on inexpensive substrates.
When benchmarked against the broader single-cell protein (SCP) landscape, the 58% protein content and 33–42 g/L biomass titers obtained here place both Y. lipolytica wild strains at the upper end of current industrial expectations. A recent review of commercial SCPs reports typical protein fractions of 45–55% dry weight, with only a few processes surpassing 60% when costly nutrient inputs are used [61]. Likewise, a survey of sustainable SCP production from non-grain feedstocks notes that yields above 50% protein remain challenging, particularly when inexpensive carbon sources such as crude glycerol are employed [62]. Against these reference ranges, the performance of NRRL Y-50997 achieved on glycerol- and glucose-based media without genetic engineering underscores their competitive potential for cost-effective, high-protein SCP manufacturing.
Moreover, both strains presented amino acid, lipid, and ash compositions that differed from each other. These findings underscore the importance of conducting detailed investigations on individual Y. lipolytica strains to maximize their potential for various uses, including their use as nutritional yeasts. These findings underscore the significant variations in chemical composition observed among the yeast strains under investigation, suggesting potential differences in their metabolic capacity and the application of these strains in various biotechnological processes. For example, it highlights the potential of strain NRRL Y-50997 in the production of yeast oils as an alternative feedstock for biodiesel production, as it has previously been recognized for its remarkable ability to accumulate lipids up to 53% of DCW [33]. In contrast, the presence of protein in the biomass was favored over the presence of lipids in this study because the nitrogen source was not a limiting factor.
Amino acid composition is important for some applications of Y. lipolytica biomass, and because it can vary from strain to strain, the amino acid profile of biomass from both strains when grown on a standard medium (YPD) was investigated. Interestingly, the predominant amino acid was lysine for both strains (Table 3), which is consistent with previously published data, where lysine is the predominant amino acid but not in terms of amount, as higher amounts of lysine were obtained in strains ATCC 9773 and NRRL Y-50997 (125.2 and 106.9 mg/g protein, respectively) [41]. Recent evaluations in various contexts consistently identify lysine as the main bottleneck in the quality of plant-based proteins; in this regard, it has been reported that in a cohort of 193 New Zealand vegans lysine intake barely met requirements, being less than adequate for approximately half of the participants after adjustment for actual ileal digestibility [63], whereas dietary modeling in the Netherlands revealed that substitution of meat with plant analogs further reduced the amount of usable lysine, making it the first indispensable amino acid in the national diet [64]. Therefore, the fact that both Y. lipolytica strains studied here accumulate lysine as the most abundant amino acid is nutritionally relevant; incorporating even modest proportions of this high-lysine biomass could correct the lysine deficit of plant-based diets and improve protein quality without resorting to synthetic amino-acid supplementation.
Additionally, both strains exhibited similar patterns in that the least abundant amino acid was cysteine, with a concentration of <1.2 mg/g protein. The data presented here are consistent with those reported in other studies where another strain, isolated from soil and designated S6, was grown with crude glycerol [58]. In that study, the strain was distinguished by having 8.26 g per 100 g of lysine protein and 5.39 g per 100 g of threonine protein, which are the amino acids with the highest levels, whereas the contents of methionine and cysteine (sulfur-containing amino acids) present in the biomass of the strain were lower. Strain ATCC 9773 exceeds the requirements for almost all FAO adult amino acids, but the amino acid levels of NRRL Y-50997 fall below the FAO requirements for isoleucine, leucine, and valine. Nevertheless, ATCC 9773 and NRRL Y-50997 are excellent alternatives to other protein sources, such as egg and cow milk, or other yeasts, such as C. utilis and S. cerevisiae.
The digestibility of Y. lipolytica is similar to that of Saccharomyces [65]; thus, Y. lipolytica can be considered a nutritional yeast, and it has been successfully applied in animal feed [41]. However, even though the safety of Y. lipolytica has been widely demonstrated [10], in some legislations, such as in Europe, it is only approved as an ingredient in human food supplements [20]. Therefore, more research is needed in this regard to enable its application in food and nutraceutical applications. Future research could evaluate functional properties of the strains studied in this work, such as immunomodulatory or antioxidant effects in animal feeding trials, by evaluating bioactive components such as β-glucans naturally present in Y. lipolytica cell walls [42,66,67]. These efforts will help to translate the nutritional and functional potential of strains NRRL Y-50997 and ATCC 9773 into validated feed applications and health-beneficial formulations.
Some of the animal species in which the efficiency of Y. lipolytica biomass has been demonstrated include turkeys, piglets, fish, shrimp, and calves (Figure 3). The reported benefits are immunostimulants, the modulation of lipid metabolism, the stimulation of the antioxidant system, the reduction in the percentage of abdominal fat content and the improvement of metabolism in turkeys [42,43,44]; improved growth performance, greater weight gain, the modulation of lipid metabolism, immunostimulants, and favorable effects on the gut microbiota in pigs [45,46]; weight gain, a greater ratio of eicosapentaenoic acid/docosahexaenoic acid, high retention of docosapentaenoic acid and docosahexaenoic acid, the modulation of gut microbial communities, and potential immunostimulants in fishes [30,47]; increased monounsaturated and polyunsaturated fatty acid contents in shrimps and immunostimulants; a higher specific growth rate; greater weight gain [48,49,50]; improved reticulorumen function, enhanced starter feed intake, better metabolism, and good growth performance during the preweaning period in calves [51].
Despite the promising results observed thus far, the widespread adoption of Y. lipolytica biomass in animal feed still faces several challenges. These include the need for further research to understand the long-term effects more fully on animal and human health, the pursuit of strains such as NRRL Y-50997, which are more efficacious for these purposes, and the optimization of production processes to ensure economic viability and environmental sustainability. Although more than 100 products have been obtained from Y. lipolytica, and it has the advantages of high-cell-density cultures and easy cell separation, downstream processing has been pointed out as an ongoing challenge in an industry survey [9]. Furthermore, regulatory acceptance and consumer awareness can act as significant barriers to the commercialization and adoption of this technology. While this work focuses on two wild-type isolates, future studies will benefit from a broader panel, including evolved and engineered strains, to capture the full genetic and metabolic diversity of Y. lipolytica. Despite these challenges, the increasing interest in the use of Y. lipolytica in animal feed suggests its significant potential to improve the efficiency and sustainability of animal production while promoting greater circularity in the biobased economy.

5. Conclusions

These findings demonstrate that Y. lipolytica strains ATCC 9773 and NRRL Y-50997 exhibit distinct metabolic profiles, with NRRL Y-50997 showing a marked preference for glycerol over glucose and promising traits for sustainable bioprocesses. Despite similarities in protein content, differences in amino acid, lipid, and ash composition highlight the importance of strain-specific characterization.
Finally, there are several promising prospects for the use of Y. lipolytica biomass in multiple applications. Some examples of these applications include bioremediation, biofuel production, and the development of functional foods. Furthermore, Y. lipolytica has the potential to serve as a sustainable source of protein and lipids for animal feed, which could address challenges related to food security and the growing demand for animal protein. Moreover, the capacity of Y. lipolytica to generate high-value-added compounds, including fatty acids, citric acid, and enzymes, is anticipated to profoundly influence biotechnology. Consequently, with continued research and refinement of metabolic engineering techniques, it is likely that current processes, where biomass production and nutritional profiles play important roles, will be optimized.

Author Contributions

Conceptualization, D.T.-A. and G.S.; methodology, D.T.-A. and G.S.; validation, D.T.-A. and G.S.; formal analysis, D.T.-A. and G.S.; investigation, D.T.-A. and G.S.; resources, D.T.-A. and G.S.; data curation, D.T.-A.; writing—original draft preparation, D.T.-A.; writing—review and editing, D.T.-A. and G.S.; visualization, D.T.-A.; supervision, G.S.; project administration, G.S. All authors have read and agreed to the published version of the manuscript.

Funding

The experimental work was conducted at the Laboratorio de Innovación en Bioenergéticos y Bioprocesos Avanzados (LIBBA), Biotechnology Unit, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ).

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author(s).

Acknowledgments

We are grateful to Secretaría de Ciencia, Humanidades, Tecnología e Innovación (Secihti), previously Consejo Nacional de Humanidades, Ciencias y Tecnologías (CONAHCYT) for the doctoral grant with CVU number 625756 (David Torres-Añorve) and to the Laboratorio de Innovación en Bioenergéticos y Bioprocesos Avanzados (LIBBA) Laboratory of the Biotechnology Unit of Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), where this work was carried out.

Conflicts of Interest

The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Abbreviations

The following abbreviations are used in this manuscript:
AAAAromatic amino acid (tryptophan + phenylalanine + tyrosine)
DHADocosahex-aenoic acid
DPADocosapentaenoic acid
EPAEicosapentaenoic acid
FAFatty acids
SSASulfur amino acid (cysteine + methionine)

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Applmicrobiol 05 00077 g001
Figure 1. Cellular growth of ATCC 9773 during flask cultivation in YPD (circle), YPD6 (triangle), YPG (square), and YPG6 (diamond) media. The ATCC 9773 strain was cultured for 72 h in YPD medium. The values shown represent the means of independent experiments performed in triplicate.
Figure 1. Cellular growth of ATCC 9773 during flask cultivation in YPD (circle), YPD6 (triangle), YPG (square), and YPG6 (diamond) media. The ATCC 9773 strain was cultured for 72 h in YPD medium. The values shown represent the means of independent experiments performed in triplicate.
Applmicrobiol 05 00077 g001
Applmicrobiol 05 00077 g002
Figure 2. Cellular growth of NRRL Y-50997 during flask cultivation in YPD (circle), YPD6 (triangle), YPG (square), and YPG6 (diamond) media. The NRRL Y-50997 strain was cultured for 72 h in YPD medium. The values shown represent the means of independent experiments performed in triplicate.
Figure 2. Cellular growth of NRRL Y-50997 during flask cultivation in YPD (circle), YPD6 (triangle), YPG (square), and YPG6 (diamond) media. The NRRL Y-50997 strain was cultured for 72 h in YPD medium. The values shown represent the means of independent experiments performed in triplicate.
Applmicrobiol 05 00077 g002
Applmicrobiol 05 00077 g003
Figure 3. Nutritional benefits of the use of Y. lipolytica biomass in turkeys [42,43,44], pigs [45,46], fishes [30,47], shrimps [48,49,50], and calves [51]. FA: Fatty acids; EPA: Eicosapentaenoic acid; DPA: Docosapentaenoic acid; DHA: Docosahexaenoic acid.
Figure 3. Nutritional benefits of the use of Y. lipolytica biomass in turkeys [42,43,44], pigs [45,46], fishes [30,47], shrimps [48,49,50], and calves [51]. FA: Fatty acids; EPA: Eicosapentaenoic acid; DPA: Docosapentaenoic acid; DHA: Docosahexaenoic acid.
Applmicrobiol 05 00077 g003
Table 1. Maximum biomass, biomass yield, and specific growth rate in flask cultures.
Table 1. Maximum biomass, biomass yield, and specific growth rate in flask cultures.
ParameterATCC 9773NRRL Y-50997
YPDYPD6YPGYPG6YPDYPD6YPGYPG6
Biomass
(DCW, g/L)		12.1 ± 0.5 a35.5 ± 0.820.6 ± 0.5 b34.2 ± 1.2 c16.5 ± 0.5 a34.5 ± 0.533.3 ± 0.6 b42 ± 0.8 c
YX/S
(g biomass/g carbon source)		0.61 ± 0.025 d0.59 ± 0.0131.03 ± 0.025 e0.57 ± 0.02 f0.83 ± 0.025 d0.58 ± 0.0081.67 ± 0.03 e0.7 ± 0.013 f
μ (h−1)0.07 ± 0060.06 ± 0.0060.08 ± 0.01 g0.05 ± 0.0030.1 ± 0.020.06 ± 0.0070.15 ± 0.02 g0.06 ± 0.007
μ: specific growth rate; YX/S: biomass to substrate yield. Superscript letters indicate differences between strains at p < 0.05 (ANOVA, n = 3).
Table 2. Biomass composition of Y. lipolytica strains.
Table 2. Biomass composition of Y. lipolytica strains.
ParameterATCC 9773 *NRRL Y-50997 *
Protein (%)58.858.2
Essential amino acids (%)62.6 a41.5 a
Lipids (%)3.046.35
Carbohydrates (%)32.3228.57
Ashes (%)5.806.88
* Percentage by weight. a for individual composition of essential amino acids, please refer to Table 3.
Table 3. Amino acid composition of Y. lipolytica strains compared with the average essential amino acid content of other protein sources (mg/g protein) and FAO requirements for adults. Table modified from Ref. [41].
Table 3. Amino acid composition of Y. lipolytica strains compared with the average essential amino acid content of other protein sources (mg/g protein) and FAO requirements for adults. Table modified from Ref. [41].
Amino AcidFAO
Requirements		ATCC 9773NRRL Y-50997Y. lipolyticaS. cerevisiaeC. utilisEggCow Milk
Arg-17.6 ± 0.1940.5 ± 1.504846.53211.533
His *1572.7 ± 0.9419.3 ± 0.372623.516437
Ile *3041.1 ± 1.8722.5 ± 0.194437486840
Leu *5997.8 ± 1.8745.0 ± 1.506863719088
Lys *45125.1 ± 2.62 a106.9 ± 2.62 b7065516378
Cys-<1.2 ± 0.00<1.2 ± 0.0011924249
Met *-19.1 ± 0.1914.6 ± 0.00121415.53229
SAA2220.3 ± 0.1915.8 ± 0.00232339.55638
Phe *-72.7 ± 0.1964.8 ± 0.374033416347
Trp *-47.9 ± 0.5641.7 ± 0.374793916ND
Tyr-17.6 ± 0.3713.2 ± 0.0066262019516
AAA38138.2 ± 1.12119.7 ± 0.741536810098.563
Thr *2335.3 ± 0.7530.4 ± 0.944848415048.7
Val *3958.8 ± 0.5638.2 ± 0.945353557447.9
* Essential amino acids; a ATCC 9773 major amino acid; b NRRL Y-50997 major amino acid; SAA: sulfur amino acid (cysteine + methionine); AAA: aromatic amino acid (tryptophan + phenylalanine + tyrosine); ND: not determined.
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Torres-Añorve, D.; Sandoval, G. Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica. Appl. Microbiol. 2025, 5, 77. https://doi.org/10.3390/applmicrobiol5030077

AMA Style

Torres-Añorve D, Sandoval G. Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica. Applied Microbiology. 2025; 5(3):77. https://doi.org/10.3390/applmicrobiol5030077

Chicago/Turabian Style

Torres-Añorve, David, and Georgina Sandoval. 2025. "Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica" Applied Microbiology 5, no. 3: 77. https://doi.org/10.3390/applmicrobiol5030077

APA Style

Torres-Añorve, D., & Sandoval, G. (2025). Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica. Applied Microbiology, 5(3), 77. https://doi.org/10.3390/applmicrobiol5030077

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