Xylitol fermentation characteristics with a newly isolated yeast Wickerhamomyces anomalus WA.

Xylitol is an increasingly popular functional food additive, and the newly isolated yeast Wickerhamomyces anomalus WA has shown extensive substrate utilization capability, with the ability to grow on hexose (d-galactose, d-glucose, d-mannose, l-fructose, and d-sorbose) and pentose (d-xylose and l-arabinose) substrates, as well as high tolerance to xylose at concentrations of up to 300 g/L. Optimal xylitol fermentation conditions were achieved at 32 °C, 140 rpm, pH 5.0, and initial cell concentration OD600 of 2.0, with YP (yeast extract 10 g/L, peptone 20 g/L) as the optimal nitrogen source. Xylitol yield increased from 0.61 g/g to 0.91 g/g with an increase in initial substrate concentration from 20 g/L to 180 g/L. Additionally, 20 g/L glycerol was found to be the optimal co-substrate for xylitol fermentation, resulting in an increase in xylitol yield from 0.82 g/g to 0.94 g/g at 140 rpm, enabling complete conversion of xylose to xylitol.

Enzymatic biotransformation of Rb3 from the leaves of Panax notoginseng to ginsenoside rd by a recombinant ß-xylosidase from Thermoascus aurantiacus.

Given the important pharmacological activity of ginsenoside Rd but its low content in plants, the production of Rd by enzymatic transformation is of interest. In this study, a ß-xylosidase gene Ta-XylQS from Thermoascus aurantiacus was cloned and overexpressed in Komagataella phaffii. Purified recombinant Ta-XylQS specifically hydrolyzes substrates with xylosyl residues at the optimal pH of 3.5 and temperature of 60 °C. This study established a process for producing Rd by transforming ginsenoside Rb3 in the saponins of Panax notoginseng leaves via recombinant Ta-XylQS. After 60 h, 3 g L- 1 of Rb3 was transformed into 1.46 g L- 1 of Rd, and the maximum yield of Rd reached 4.31 g kg- 1 of Panax notoginseng leaves. This study is the first report of the biotransformation of ginsenoside Rb3 to Rd via a ß-xylosidase, and the established process could potentially be adopted for the commercial production of Rd from Rb3.

The clinical characteristics of Hirayama disease in females.

INTRODUCTION: To characterize Hirayama disease in female patients, and increase awareness among clinicians regarding the specifics of this disease. METHODS: Baseline data, clinical manifestations, characteristics of cervical-flexion magnetic resonance imaging, and electromyography were collected and compared among females and males with Hirayama disease. In addition, the literature on Hirayama disease in females up to October, 2021 was searched in PubMed and the relevant data were compared with the data from our study. RESULTS: Twenty female and 40 male patients were included in this study. The average ages of onset and menarche were 14.65 and 12.75 years old. All patients suffered from muscular weakness and atrophy of the upper limb(s), with flattening and/or atrophy of the lower cervical spinal cords in cervical-flexion magnetic resonance imaging, and neurogenic patterns in the atrophic muscles as determined using electromyography. The age of onset in females was about 2 years later than the age of menarche, and the age of onset in females was 2 years earlier than that in males. There were no obvious differences in clinical presentation between males and females. DISCUSSION: Although females presented with Hirayama disease two years earlier than males, no other clinical differences were observed. Hirayama disease is likely associated with growth and development in puberty, and early identification, regardless of whether patients are male or female, is critical to optimizing prognosis.

F-waves of peroneal and tibial nerves in the differential diagnosis and follow-up evaluation of L5 and S1 radiculopathies.

PURPOSE: To investigate F-wave as a method to identify a specific root lesion of L5 or S1 and to quantitatively evaluate the severity and progression of motor root lesions in lumbosacral radiculopathies (LR). METHODS: Both peroneal and tibial F-waves were performed bilaterally in 142 patients with unilateral L5 or S1 radiculopathies and 37 controls along with Medical Research Council (MRC) evaluation, and soleus H-reflexes were tested bilaterally in 78 of these 142 cases. Both F-wave and MRC were re-evaluated approximately 1 year after initial examination in 65 patients. RESULTS: Abnormal peroneal F-waves were found in 34 patients with L5 radiculopathy (34/67, 50.7%) along with normal tibial F-waves and soleus H-reflexes in all tested cases. By contrast, 27 patients with S1 radiculopathy presented abnormal tibial F-waves (27/76, 36.0%) along with normal peroneal F-waves in all 76 cases and abnormal soleus H-reflexes in 38 of 47 (80.9%) cases. There were significant differences in involved side F-duration among different MRC scales in both radiculopathy groups (P < 0.05). Follow-up analysis demonstrated slow progression of both F-wave abnormalities and muscle weakness in patients undergoing conservative treatment (P < 0.05). CONCLUSIONS: Comparisons of F-waves between the same nerve on both sides and between peroneal and tibial nerves in the same leg may clearly increase the validity of F-waves for evaluating a specific motor root lesion of L5 or S1. Furthermore, a quantitative comparison of F-waves may provide additional information on the severity of individual root lesions and their progression even in the early stages of disease. These slides can be retrieved under Electronic Supplementary Material.

Psychometric evaluation of the Chinese version of the revised American Pain Society Patient Outcome Questionnaire concerning pain management in Chinese orthopedic patients.

The present study tested the clinical efficiency (item grouping, internal consistency of the subscales, construct validity, and clinical feasibility) of a widely used pain assessment system, the Mandarin version of the American Pain Society Patient Outcome Questionnaire (APS-POQ-R-C), in Chinese patients. We also attempted to investigate the current quality of pain management provided in orthopedic inpatient units in China and provide baseline data. First, we investigated the test-retest reliability of APS-POQ-R-C. In total, 236 orthopedic patients were evaluated. Our results showed that APS-POQ-R-C has satisfactory internal consistency and construct validity, although some items are not appropriate for orthopedic patients. Test-retest reliability outcomes indicated that APS-POQ-R-C is a satisfactory battery with acceptable validity and reliability, and is therefore recommended for pain management in future studies.

Butanol production employing fed-batch fermentation by Clostridium acetobutylicum GX01 using alkali-pretreated sugarcane bagasse hydrolysed by enzymes from Thermoascus aurantiacus QS 7-2-4.

Sugarcane bagasse (SB) is a potential feedstock for butanol production. However, biological production of butanol from SB is less economically viable. In this study, evaluation of eight pretreatments on SB showed that alkali pretreatment efficiently removed lignin from SB while retaining the intact native structure of the released microfibrils. In total, 99% of cellulose and 100% of hemicellulose in alkali-pretreated SB were hydrolysed by enzymes from Thermoascus aurantiacus. The hydrolysate was used to produce butanol in a fed-batch fermentation by Clostridium acetobutylicum. At 60h, 14.17 and 21.11gL(-1) of butanol and acetone-butanol-ethanol (ABE) were produced from 68.89gL(-1) of total sugars, respectively, yielding 0.22 and 0.33gg(-1) of sugars. The maximum yield of butanol and ABE reached 15.4g and 22.9g per 100g raw SB, respectively. This established process may have potential application for butanol production from SB.

Alcohol dehydrogenases from Kluyveromyces marxianus: heterologous expression in Escherichia coli and biochemical characterization.

BACKGROUND: Kluyveromyces marxianus has recently become a species of interest for ethanol production since it can produce ethanol at high temperature and on a wide variety of substrates. However, the reason why this yeast can produce ethanol at high temperature is largely unknown. RESULTS: The ethanol fermentation capability of K. marxianus GX-UN120 at 40°Ð¡ was found to be the same as that of Saccharomyces cerevisiae at 34°Ð¡. Zymogram analysis showed that alcohol dehydrogenase 1 (KmAdh1) was largely induced during ethanol production, KmAdh4 was constitutively expressed at a lower level and KmAdh2 and KmAdh3 were almost undetectable. The genes encoding the four alcohol dehydrogenases (ADHs) were cloned from strain GX-UN120. Each KmADH was expressed in Escherichia coli and each recombinant protein was digested with enterokinase to remove the fusion protein. The optimum pH of the purified recombinant KmAdh1 was 8.0 and that of KmAdh2, KmAdh3 and KmAdh4 was 7.0. The optimum temperatures of KmAdh1, KmAdh2, KmAdh3 and KmAdh4 were 50, 45, 55 and 45°C, respectively. The K(m) values of the recombinant KmAdh1 and KmAdh2 were 4.0 and 1.2 mM for acetaldehyde and 39.7 and 49.5 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh1 and KmAdh2 were 114.9 and 21.6 µmol min⁻¹ mg⁻¹ for acetaldehyde and 57.5 and 1.8 µmol min⁻¹ mg⁻¹ for ethanol, respectively. KmAdh3 and KmAdh4 catalyze the oxidation reaction of ethanol to acetaldehyde but not the reduction reaction of acetaldehyde to ethanol, and the K(m) values of the recombinant KmAdh3 and KmAdh4 were 26.0 and 17.0 mM for ethanol, respectively. The V(max) values of the recombinant KmAdh3 and KmAdh4 were 12.8 and 56.2 µmol min⁻¹ mg⁻¹ for ethanol, respectively. CONCLUSION: These data in this study collectively indicate that KmAdh1 is the primary ADH responsible for the production of ethanol from the reduction of acetaldehyde in K. marxianus. The relatively high optimum temperature of KmAdh1 may partially explain the ability of K. marxianus to produce ethanol at high temperature. Understanding the biochemical characteristics of KmAdhs will enhance our fundamental knowledge of the metabolism of ethanol fermentation in K. marxianus.

A new analytical potential energy surface for the singlet state of He2H+.

The analytic potential energy surface (APES) for the exchange reaction of HeH(+) (X(1)Σ(+)) + He at the lowest singlet state 1(1)A(∕) has been built. The APES is expressed as Aguado-Paniagua function based on the many-body expansion. Using the adaptive non-linear least-squares algorithm, the APES is fitted from 15 682 ab initio energy points calculated with the multireference configuration interaction calculation with a large d-aug-cc-pV5Z basis set. To testify the new APES, we calculate the integral cross sections for He + H(+)He (v = 0, 1, 2, j = 0) → HeH(+) + He by means of quasi-classical trajectory and compare them with the previous result in literature.

Fermentation of xylose into ethanol by a new fungus strain Pestalotiopsis sp. XE-1.

A new fungus, Pestalotiopsis sp. XE-1, which produced ethanol from xylose with yield of 0.47 g ethanol/g of consumed xylose was isolated. It also produced ethanol from arabinose, glucose, fructose, mannose, galactose, cellobiose, maltose, and sucrose with yields of 0.38, 0.47, 0.45, 0.46, 0.31, 0.25, 0.31, and 0.34 g ethanol/g of sugar consumed, respectively. It produced maximum ethanol from xylose at pH 6.5, 30°C under a semi-aerobic condition. Acetic acid produced in xylose fermenting process inhibited ethanol production of XE-1. The ethanol yield in the pH-uncontrolled batch fermentation was about 27% lower than that in the pH-controlled one. The ethanol tolerance of XE-1 was higher than most xylose-fermenting, ethanol-producing microbes, but lower than Saccharomyces cerevisiae and Hansenula polymorpha. XE-1 showed tolerance to high concentration of xylose, and was able to grow and produce ethanol even when it was cultivated in 97.71 g/l xylose.

Multiple induced mutagenesis for improvement of ethanol production by Kluyveromyces marxianus.

Kluyveromyces marxianus GX-15 was mutated multiple times by alternately treatment with UV irradiation and NTG for two cycles. Four mutant strains with improved ethanol yield were obtained. The maximum ethanol concentration, ethanol yield coefficient and theoretical ethanol yield of the best mutant strain, GX-UN120, was 69 g/l, 0.46 g/g and 91%, respectively, when fermenting 150 g glucose/l at 40°C. The corresponding values for GX-15 were 58 g/l, 0.39 g/g and 76%, respectively. GX-UN120 grew well in 11% (v/v) of ethanol, while GX-15 could not grow when ethanol was greater than 8% (v/v).