Comparative transcriptome analysis reveals the redirection of metabolic flux from cell growth to astaxanthin biosynthesis in Yarrowia lipolytica.

Engineering Yarrowia lipolytica to produce astaxanthin provides a promising route. Here, Y. lipolytica M2 producing a titer of 181 mg/L astaxanthin was isolated by iterative atmospheric and room-temperature plasma mutagenesis and diphenylamine-mediated screening. Interestingly, a negative correlation was observed between cell biomass and astaxanthin production. To reveal the underlying mechanism, RNA-seq analysis of transcriptional changes was performed in high producer M2 and reference strain M1, and a total of 1379 differentially expressed genes were obtained. Data analysis revealed that carbon flux was elevated through lipid metabolism, acetyl-CoA and mevalonate supply, but restrained through central carbon metabolism in strain M2. Moreover, upregulation of other pathways such as ATP-binding cassette transporter and thiamine pyrophosphate possibly provided more cofactors for carotenoid hydroxylase and relieved cell membrane stress caused by astaxanthin insertion. These results suggest that balancing cell growth and astaxanthin production may be important to promote efficient biosynthesis of astaxanthin in Y. lipolytica.

Association of fat distribution differences in infertile women with assisted reproductive outcomes: A prospective cohort study.

OBJECTIVE: To investigate the impacts of fat distribution on assisted reproductive outcomes in infertile women. METHODS: The study randomly recruited 576 infertile women who underwent assisted reproductive technology treatment at the Reproductive Medicine Center of the First Affiliated Hospital of Anhui Medical University between July and October 2022. Questionnaires and body composition measurements were administered to assess baseline information and fat distribution. The numbers of oocytes, zygotes presenting with two pronuclei (2PN), and available embryos were tracked at the end of the cycle. Multifactorial logistic regression models and restricted cubic spline (RCS) curve models were used to explore the relationships between fat distribution and reproductive outcomes while controlling for confounding factors. RESULTS: The study found that the participants had a mean age of 30.82 years. The analysis showed that there was a significant difference between the amount of leg body fat mass (LBFM) and the distribution of reproductive outcomes. However, there was no significant correlation between the level of visceral fat and reproductive outcomes. After taking confounding factors into account, the multifactorial regression analysis showed that the total body fat mass and the number of oocytes (odds ratio (OR) 0.92, 95% confidence interval (CI) 0.84-0.99), 2PN (OR 0.92, 95% CI 0.84-0.99), and embryos available for transfer (OR 0.90, 95% CI 0.82-0.99) were negatively correlated. RCS modeling revealed a linear dose-response relationship between LBFM and assisted reproductive outcomes. CONCLUSION: Fat distribution varies among infertile women, and higher amounts of fat are associated with poorer assisted reproductive outcomes.

A study on the impact of the number of family generations on intergenerational support for centenarians: a study in a Chinese 'longevity city'.

BACKGROUND: As a natural source of support for the elderly, the family is an important channel for achieving a sense of security, happiness, and worthiness in old age. In this study, we analysed the characteristics of intergenerational support in families of centenarians and explored the impact of the number of family generations on intergenerational support. METHODS: We conducted a cross-sectional survey between April 2020 and January 2021 among 62 elderly people aged 99+ in Rugao, China, one of six 'longevity cities' in the world. Assisted by the researchers, centenarians completed questionnaires with details pertaining to general demographics, intergenerational support, and other aspects. We used a logistic regression model to analyse the influence of the number of family generations on intergenerational support that the centenarians received with respect to economic, living, and emotional aspects. RESULTS: Centenarians were primarily recipients of care in their families, and received intergenerational support mainly for their declined physical functions and limited self-care ability. The study results revealed that the greater the number of generations comprising the family, the greater was the intergenerational life care and emotional comfort provided for centenarians by the family. CONCLUSIONS: In this study, we found a positive effect of the number of family generations on intergenerational support for centenarians. The government and society should promote the tradition of respecting, caring for, and honouring the elderly while paying close attention to the dynamic changes in the family structure of centenarians in promoting high-quality and sustainable development of the people, economy, and society.

Bacterial infection promotes tumorigenesis of colorectal cancer via regulating CDC42 acetylation.

Increasing evidence highlights the role of bacteria in promoting tumorigenesis. The underlying mechanisms may be diverse and remain poorly understood. Here, we report that Salmonella infection leads to extensive de/acetylation changes in host cell proteins. The acetylation of mammalian cell division cycle 42 (CDC42), a member of the Rho family of GTPases involved in many crucial signaling pathways in cancer cells, is drastically reduced after bacterial infection. CDC42 is deacetylated by SIRT2 and acetylated by p300/CBP. Non-acetylated CDC42 at lysine 153 shows an impaired binding of its downstream effector PAK4 and an attenuated phosphorylation of p38 and JNK, consequently reduces cell apoptosis. The reduction in K153 acetylation also enhances the migration and invasion ability of colon cancer cells. The low level of K153 acetylation in patients with colorectal cancer (CRC) predicts a poor prognosis. Taken together, our findings suggest a new mechanism of bacterial infection-induced promotion of colorectal tumorigenesis by modulation of the CDC42-PAK axis through manipulation of CDC42 acetylation.

Resilience of infertile families undergoing in vitro fertilization: An application of the double ABC-X model.

BACKGROUND: Family resilience plays a crucial role in protecting the mental health and family stability of infertile patients. However, information associated with infertile families resilience is scarce. The double ABC-X model provides a roadmap for this, helps organize knowledge, and lays the foundation for knowledge development. AIMS: To describe the current situation of family resilience of infertile women, and to test the predictive theoretical model of family resilience based on infertility stigma, individual resilience, coping style, and posttraumatic growth. DESIGN: A cross-sectional study. METHODS: A convenience sample of 372 infertile women undergoing in vitro fertilization were recruited between April and August 2020. The Chinese-Family Resilience Assessment Scale, Infertility Stigma Scale, Simplified Coping Style Questionnaire, Chinese version of Connor-Davidson Resilience Scale, and Chinese version of Post Traumatic Growth Inventory were used to measure family resilience, infertility stigma, individual resilience, coping style, and posttraumatic growth. Structural equation models were used to analyze the relationship among these variables. RESULTS: The results showed that family resilience was related to infertility stigma, positive coping, and individual resilience. Moreover, the path analysis indicated that positive coping and individual resilience mediated the effects of infertility stigma on family resilience. CONCLUSIONS: A high level of stigma among infertile women should be identified. Interventions for targeting positive coping and individual resilience might ultimately increase their family resilience.

Pancreatic paraganglioma with multiple lymph node metastases found by spectral computed tomography: A case report and review of the literature.

BACKGROUND: Primary pancreatic paraganglioma is exceedingly rare. Most patients with pancreatic paraganglioma lack a typical clinical presentation, and the tumor is difficult to accurately differentiate from other pancreatic neuroendocrine tumors, making the misdiagnosis rate extremely high. Surgical excision is the primary treatment modality but is considered high risk. Because of its rich vascularity, the tumor easily bleeds during surgery, especially malignant paragangliomas invading large blood vessels. Thus, a thorough preoperative evaluation of the tumor is necessary. Here, we report a primary malignant pancreatic paraganglioma, the second such case in a young patient that was successfully resected surgically. CASE SUMMARY: A 26-year-old female patient was admitted to the hospital with unexplained abdominal pain. Dual-layer spectral-detector computed tomography (DLCT) revealed a mixed density mass in the pancreatic body and tail. The patient was transferred to our hospital after previous failed surgical resection at other hospitals. The patient and her family strongly desired surgery. After a thorough preoperative evaluation and adequate preparation, a large mass with the greatest dimension of 8.0 cm was successfully resected. The final pathological diagnosis was malignant paraganglioma. The patient was discharged in good condition 2 wk postoperatively. CONCLUSION: The rare malignant pancreatic paraganglioma reported here was difficult to diagnose preoperatively. Early filling of the draining vein may be a crucial diagnostic imaging feature. DLCT can provide more precise information for surgical resection through dual-energy imaging.

Single-Cell Transcriptomics Reveals Longevity Immune Remodeling Features Shared by Centenarians and Their Offspring.

Centenarians, who show mild infections and low incidence of tumors, are the optimal model to investigate healthy aging. However, longevity related immune characteristics has not been fully revealed largely due to lack of appropriate controls. In this study, single-cell transcriptomic analysis of peripheral blood mononuclear cells (PBMCs) derived from seven centenarians (CEN), six centenarians' offspring (CO), and nine offspring spouses or neighbors (Control, age-matched to CO) are performed to investigate the shared immune features between CEN and CO. The results indicate that among all 12 T cell clusters, the cytotoxic-phenotype-clusters (CPC) and the naïve-phenotype-clusters (NPC) significantly change between CEN and ontrol. Compared to Control, both CEN and CO are characterized by depleted NPC and increased CPC, which is dominated by CD8+ T cells. Furthermore, CPC from CEN and CO share enhanced signaling pathways and transcriptional factors associated with immune response, and possesse similar T-cell-receptor features, such as high clonal expansion. Interestingly, rather than a significant increase in GZMK+ CD8 cells during aging, centenarians show accumulation of GZMB+ and CMC1+ CD8 T cells. Collectively, this study unveils an immune remodeling pattern reflected by both quantitative increase and functional reinforcement of cytotoxic T cells which are essential for healthy aging.

Sexual Function, Self-Esteem, and Quality of Life in Infertile Couples Undergoing in vitro Fertilization: A Dyadic Approach.

Purpose: Infertility has adverse effects on the quality of life (QoL) of infertile couples. Previous studies found important associations between sexual function, self-esteem and QoL, but mainly focused on one individual's approach rather than both partners. This study adopted a dyadic approach to evaluate the relationship between sexual function and QoL in couples with infertility through mediation and improving self-esteem. Patients and Methods: Between October 2020 and January 2021, 428 couples with infertility (n=856) undergoing in-vitro fertilization (IVF) at a tertiary hospital in Hefei, China, were registered for the current descriptive cross-sectional research. The dyads' sociodemographic and clinical features, as well as their sexual function, self-esteem, and QoL were evaluated. The Fertility quality of life scale (FertiQoL), Rosenberg Self-Esteem Scale (RSES), Female Sexual Function Index (FSFI), and International Index of Erectile Function-15 (IIEF-15) were used to evaluate the participants. The Actor-Partner Interdependence Mediation Model (APIMeM) was utilized to examine data from the dyadic relationships. Results: According to the APIMeM analysis, sexual function of individuals with infertility was directly and indirectly connected with their QoL, mediated through their self-esteem. The women's sexual function was found to be positively associated with their partner's QoL, with the women's self-esteem acting as a complete mediator. The men's sexual function was found to be positively associated with partner's QoL, with the men's self-esteem acting as a complete mediator. Conclusion: The findings suggest that boosting participants' self-esteem can help them and their partners have a better QoL. Also, therapies aimed at improving and sustaining self-esteem of couples with infertility could help mitigate the negative influence of low sexual function on their QoL.

[Quality evaluation of Psoraleae Fructus based on QAMS method and TLC identification].

The present study established a determination method of Psoraleae Fructus by quantitative analysis of multi-components by the single marker(QAMS) and further improved the thin-layer chromatography(TLC) method. The QAMS method was established by UPLC with psoralen as the internal marker, and the content of psoralenoside, isopsoralenoside, psoralen, and isopsoralen was simultaneously determined. As revealed by the comparison with results of the external standard method, the QAMS method was accurate and feasible. According to the current quality standards of Psoraleae Fructus, the TLC method was further optimized and improved, and bakuchiol was added for identification based on the original TLC method with psoralen and isopsoralen as indicators. This study provides a reference for improving the quality control method of Psoraleae Fructus.

Integrated pathway engineering and transcriptome analysis for improved astaxanthin biosynthesis in Yarrowia lipolytica.

Astaxanthin is a high value carotenoid with a broad range of commercial applications due to its superior antioxidant properties. In this study, ß-carotene-producing Yarrowia lipolytica XK17 constructed in the lab was employed for astaxanthin biosynthesis. The catalytic effects of ß-carotene ketolase CrtW and ß-carotene hydroxylase CrtZ from various species were investigated. The PspCrtW from Paracoccus sp. and HpCrtZ# from Haematococcus pluvialis were confirmed to be the best combination in converting ß-carotene. Several key bottlenecks in biomass and astaxanthin biosynthesis were effectively eliminated by optimizing the expression of the above enzymes and restoring uracil/leucine biosynthesis. In addition, the effects of astaxanthin biosynthesis on cell metabolism were investigated by integrated analysis of pathway modification and transcriptome information. After further optimization, strain DN30 was able to synthesize up to 730.3 mg/L astaxanthin in laboratory 5-L fermenter. This study provides a good metabolic strategy and a sustainable development platform for high-value carotenoid production.

CRISPR/Cas9-based genome editing for the modification of multiple duplications that cause Duchenne muscular dystrophy.

With the development of basic research, some genetic-based methods have been found to treat Duchenne muscular dystrophy (DMD) with large deletion mutations and nonsense mutations. Appropriate therapeutic approaches for repairing multiple duplications are limited. We used the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 system with patient-derived primary myoblasts to correct multiple duplications of the dystrophin gene. Muscle tissues from a patient carrying duplications of dystrophin were obtained, and tissue-derived primary cells were cultured. Myoblasts were purified with an immunomagnetic sorting system using CD56 microbeads. After transduction by lentivirus with a designed single guide RNA (sgRNA) targeting a duplicated region, myoblasts were allowed to differentiate for 7 days. Copy number variations in the exons of the patient's myotubes were quantified by real-time PCR before and after genetic editing. Western blot analysis was performed to detect the full-length dystrophin protein before and after genetic editing. The ten sequences predicted to be the most likely off-targets were determined by Sanger sequencing. The patient carried duplications of exon 18-25, dystrophin protein expression was completely abrogated. Real-time PCR showed that the copy number of exon 25 in the patient's myotubes was 2.015 ± 0.079 compared with that of the healthy controls. After editing, the copy number of exon 25 in the patient's modified myotubes was 1.308 ± 0.083 compared with that of the healthy controls (P < 0.001). Western blot analysis revealed no expression of the dystrophin protein in the patient's myotubes before editing. After editing, the patient's myotubes expressed the full-length dystrophin protein at a level that was ~6.12% of that in the healthy control samples. Off-target analysis revealed no abnormal editing at the ten sites predicted to be the most likely off-target sites. The excision of multiple duplications by the CRISPR/Cas9 system restored the expression of full-length dystrophin. This study provides proof of evidence for future genome-editing therapy in patients with DMD caused by multiple duplication mutations.

Transcriptomic Analysis of Cadmium Stressed Tamarix hispida Revealed Novel Transcripts and the Importance of Abscisic Acid Network.

Cadmium (Cd) pollution is widely detected in soil and has been recognized as a major environmental problem. Tamarix hispida is a woody halophyte, which can form natural forest on the desert and soil with 0.5 to 1% salt content, making it an ideal plant for the research on response to abiotic stresses. However, no systematic study has investigated the molecular mechanism of Cd tolerance in T. hispida. In the study, RNA-seq technique was applied to analyze the transcriptomic changes in T. hispida treated with 150 µmol L-1 CdCl2 for 24, 48, and 72 h compared with control. In total, 72,764 unigenes exhibited similar sequences in the Non-redundant nucleic acid database (NR database), while 36.3% of all these unigenes may be new transcripts. In addition, 6,778, 8,282, and 8,601 DEGs were detected at 24, 48, and 72 h, respectively. Functional annotation analysis indicated that many genes may be involved in Cd stress response, including ion bonding, signal transduction, stress sensing, hormone responses and ROS metabolism. A ThUGT gene from the abscisic acid (ABA) signaling pathway can enhance Cd resistance ability of T. hispida by regulating the production of ROS under Cd stress and inhibit absorption of Cd. The new transcriptome resources and data that we present in this study for T. hispida may facilitate investigation of molecular mechanisms governing Cd resistance.

Health related quality of life in Chinese pregnant women at advanced maternal age: a cross-sectional study.

OBJECTIVE: This study aims to explore the relationship between advanced maternal age (AMA) and health-related quality of life in Chinese pregnant women. METHODS: A cross-sectional study was conducted in Guangzhou, China between September 2018 and June 2019. Four hundred and twenty-seven AMA women and the equal number of their younger counterparts completed the 36-Item Short-Form Health Survey (SF-36). RESULTS: Compared with younger women, the AMA women were more likely to be employed; have a higher monthly household income and insurance covered; have a satisfied relationship with their husband and mother-in-law; and had a significantly lower level of physical (SF36-PCS) health-related quality of life and a higher level of mental (SF36-MCS) health-related quality of life during the pregnancy. The association of maternal age with health-related quality of life varies according with the trimester of pregnancy. Maternal age was a significant predictor of SF36-PCS and SF36-MCS. The third trimester was the significant predictor of SF36-PCS while the relationship with the mother-in-law was the significant predictor of SF36-MCS. CONCLUSIONS: The SF36-PCS in the AMA women decreased with advancing age. However, their SF36-MCS was better over their younger counterparts. Age-related biological disadvantages may be offset by social/psychological advantages in AMA women.

Higher dietary diversity as a protective factor against depression among older adults in China: a cross-sectional study.

BACKGROUND: Evidence suggests that poor mental health (MH) is a risk factor for the health of older adults. Dietary diversity is considered to be related to healthy aging. However, the relationship between diet and MH is still unclear. Therefore, the purpose of this study was to investigate the relationship between dietary diversity score (DDS) and anxiety and depression among centenarians and their offspring and spouses. METHODS: Our study was observational and cross-sectional. The Generalized Anxiety Disorder Scale (GAD-7), the 15-item version of the Geriatric Depression Scale (GDS-15), and the dietary frequency questionnaire were used to measure the status of anxiety, depression, and dietary diversity. Data were analyzed by Student's t-test, χ2 test, Mann-Whitney U test, correlational analysis, and univariate or multivariate logistic regression. RESULTS: Among the 288 older adults, 12.8% reported symptoms of depression, and 8.7% reported anxiety. People with a lower dietary diversity had higher rates of anxiety and depression. After controlling for age, place of residence, economic status, alcohol drinking, and physical activity, a lower DDS was found to be a risk factor for depressive symptoms [odds ratio (OR): 2.237; 95% confidence interval (CI): 1.009-4.959; P=0.048]. DDS was negatively correlated with depression score in older adults (r=-0.224; P<0.001), especially offspring and their spouses (r=-0.275; P<0.001). However, no significant relationship was observed between DDS and anxiety. In addition, eating legumes (OR: 0.415; 95% CI: 0.188-0.920; P=0.030) and nuts (OR: 0.255; 95% CI: 0.116-0.561; P=0.001) at least once a week can act as protective factors for depression. Eating nuts (OR: 0.405; 95% CI: 0.168-0.978; P=0.044) and meat (OR: 0.396, 95% CI: 0.161-0.975; P=0.044) at least once a week can act as protective factors for anxiety. CONCLUSIONS: These results suggest an association between low dietary diversity and a higher incidence of mental disorders. Further, the possibility of reverse causality cannot be ruled out. It is necessary to conduct further prospective studies.

Mindfulness-based intervention in Chinese pregnant women with recurrent miscarriage: A non-randomized controlled study.

OBJECTIVE: Pregnant women with recurrent miscarriage have high rates of anxiety and depression. Mindfulness-based interventions have shown benefits in improving mental health in diverse populations; however, few studies have explored their efficacy in pregnant women with recurrent miscarriage, which was investigated in the present study. DESIGN: A nonrandomized controlled study was carried out from August 2019 to November 2020. SETTING: The study was conducted at a regional teaching hospital in Guangzhou, China that provides leading care for recurrent miscarriage. PARTICIPANTS: A total of 158 pregnant women with recurrent miscarriage were recruited and allocated to the intervention group (n = 79) or the control group (n = 79); 131 women completed the study. INTERVENTION: The mindfulness-based intervention consisted of a 1-h education session and daily mindfulness exercises guided by audio recordings during hospitalization. MEASUREMENTS AND FINDINGS: Study outcomes included perceived stress measured with the Perceived Stress Scale; symptoms of anxiety and depression measured with the Self-rating Anxiety Scale and Edinburgh Postnatal Depression Scale, respectively; and positive and negative affect measured with the Positive Affect and Negative Affect Scale - Revised, respectively. Compared to the control group, participants in the intervention group showed significant decreases in perceived stress, depression symptoms, and negative affect and an increase in positive affect after the intervention. Anxiety increased significantly in the control group during the study but remained unchanged in the intervention group. KEY CONCLUSIONS: A mindfulness-based intervention can reduce psychological symptoms and improve mental health in pregnant women with recurrent miscarriage. IMPLICATIONS FOR PRACTICE: A mindfulness-based intervention should be incorporated into routine care to help improve the mental health of pregnant women with recurrent miscarriage.

Homoharringtonine inhibited breast cancer cells growth via miR-18a-3p/AKT/mTOR signaling pathway.

Homoharringtonine (HHT), a natural alkaloid derived from the cephalotaxus, exhibited its anti-cancer effects in hematological malignancies clinically. However, its pesticide effects and mechanisms in treating solid tumors remain unclear. In this study, we found that HHT was capable of inhibiting tumor growth after 5-days treatment of breast cancer cells, MCF-7, in vivo. Furthemore, HHT also significantly inhibited the cancer cell growth and induced cell apoptosis in vitro. miRNA sequencing proved miR-18a-3p was noticeably downregulated in the cells after HHT treatment. Moreover, downregulating miR-18a-3p increased HHT-induced cell apoptosis; our data supported that HHT suppressed miR-18a-3p expression and inhibited tumorigenesis might via AKT-mTOR signaling pathway. In conclusion: our study proved that HHT suppressed breast cancer cell growth and promoted apoptosis mediated by regulating of the miR-18a-3p-AKT-mTOR signaling pathway, HHT may be a promising antitumor agent in breast cancer treatment.

A digital microfluidic platform based on a near-infrared light-responsive shape-memory micropillar array.

In this work, we developed a digital microfluidic platform based on a shape memory micropillar array responsive to near-infrared light, and the droplets were programmatically manipulated through light-induced micropillar deformation. The micropillar array was constructed on the surface of a poly(ethylene-vinyl acetate) copolymer, a shape memory polymer sensitive to near-infrared light. Before droplet manipulation, the micropillar array was kept temporarily tilted by heating and pressing. Under the irradiation of a near-infrared laser, the micropillar array achieved the transition from the temporary shape to the original shape. Temperature gradient and micropillar deformation caused by near-infrared light irradiation produce the driving force for droplet movement. The movement of the laser mounted on an electronically controlled displacement platform was controlled by a computer to achieve the programmed control of the droplets. Moreover, we demonstrated light-manipulated droplet movement and fusion, and achieved ascorbic acid detection using this digital microfluidic platform. In particular, the micropillar array chip is able to manipulate droplets in a wide range of 0.1 µL to 10 µL. The proposed digital microfluidic platform will broaden the application of digital microfluidic technology in analytical chemistry and biomedicine.

Reducing discomfort of eye drops prior to retinal examination in the neonatal intensive care unit.

OBJECTIVE: To evaluate the effectiveness in reducing pain by giving oral sucrose and non-pharmacological comfort measures prior to topical administration of mydriatic eye drops on premature infants undergoing retinopathy of prematurity (ROP) screening eye exams in a neonatal intensive care unit (NICU). STUDY DESIGN: A prospective quality improvement study was conducted in the NICU where infants were given oral sucrose prior to administration of mydriatic eye drops while a second person performed facilitated tucking and containment. Premature Infant Pain Profile (PIPP) scores were recorded during eye drop administration and compared to a group that did not receive any comfort measures. RESULT: Sixty-eight infants were enrolled. Mean PIPP scores increased an average of 1.5 (SD = 1.5) during administration of mydriatic drops without comfort measures compared to 0.6 (SD = 0.8) when comfort measures were used. This difference was statistically significant (p < 0.001). CONCLUSIONS: Oral sucrose and simple comfort measures can be effective in reducing pain associated with mydriatic eye drops.

Engineering oleaginous yeast Yarrowia lipolytica for enhanced limonene production from xylose and lignocellulosic hydrolysate.

Limonene, a valuable cyclic monoterpene, has been broadly studied in recent decades due to its wide application in the food, cosmetics and pharmaceutical industries. Engineering of the yeast Yarrowia lipolytica for fermentation of renewable biomass lignocellulosic hydrolysate may reduce the cost and improve the economics of bioconversion for the production of limonene. The aim of this study was to engineer Y. lipolytica to produce limonene from xylose and low-cost lignocellulosic feedstock. The heterologous genes XR and XDH and native gene XK encoding xylose assimilation enzymes, along with the heterologous genes tNDPS1 and tLS encoding orthogonal limonene biosynthetic enzymes, were introduced into the Po1f strain to facilitate xylose fermentation to limonene. The initially developed strain produced 0.44 mg/L of limonene in 72 h with 20 g/L of xylose. Overexpression of genes from the mevalonate pathway, including HMG1 and ERG12, significantly increased limonene production from xylose to ∼9.00 mg/L in 72 h. Furthermore, limonene production peaked at 20.57 mg/L with 50% hydrolysate after 72 h when detoxified lignocellulosic hydrolysate was used. This study is the first to report limonene production by yeast from lignocellulosic feedstock, and these results indicate the initial steps toward economical and sustainable production of isoprenoids from renewable biomass by engineered Y. lipolytica.