Screening of a Saccharomyces cerevisiae Strain with High 3-Methylthio-1-Propanol Yield and Optimization of Its Fermentation Conditions.

3-Methylthio-1-propanol (3-Met) is an important flavor compound in various alcoholic beverages such as Baijiu and Huangjiu. To maintain the content of 3-Met in these alcoholic beverages, it is necessary to screen a micro-organism with high yield of 3-Met from the brewing environment. In this study, the ability of yeast strains from the Baijiu brewing to produce 3-Met was analyzed, aiming to obtain yeast with high-yield 3-Met, and its fermentation conditions were optimized. Firstly, 39 yeast strains were screened using 3-Met conversion medium. The results showed that the majority of the strains from Baijiu brewing sources could produce 3-Met, and nearly half of the strains produced more than 0.5 g/L of 3-Met. Among these, yeast F10404, Y03401, and Y8#01, produced more than 1.0 g/L of 3-Met, with yeast Y03401 producing the highest amount at 1.30 g/L. Through morphological observation, physiological and biochemical analysis, and molecular biological identification, it was confirmed that yeast Y03401 was a Saccharomyces cerevisiae. Subsequently, the optimal fermentation conditions for 3-Met production by this yeast were obtained through single-factor designs, Plackett-Burman test, steepest ascent path design and response surface methodology. When the glucose concentration was 60 g/L, yeast extract concentration was 0.8 g/L, L-methionine concentration was 3.8 g/L, initial pH was 4, incubation time was 63 h, inoculum size was 1.6%, shaking speed was 150 rpm, loading volume was 50 mL/250 mL, and temperature was 26 °C, the content of 3-Met produced by S. cerevisiae Y03401 reached a high level of 3.66 g/L. It was also noteworthy that, in contrast to other study findings, this yeast was able to create substantial amounts of 3-Met even in the absence of L-methionine precursor. Based on the clear genome of S. cerevisiae and its characteristics in 3-Met production, S. cerevisiae Y03401 had broad prospects for application in alcoholic beverages such as Baijiu.

Optimization of High-Density Fermentation Conditions for Saccharomycopsis fibuligera Y1402 through Response Surface Analysis.

Saccharomycopsis fibuligera, which produces enzymes like amylase and protease as well as flavor substances like ß-phenyl ethanol and phenyl acetate, plays a crucial role in traditional fermented foods. However, this strain still lacks a high-density fermentation culture, which has had an impact on the strain's industrial application process. Therefore, this study investigated the optimization of medium ingredients and fermentation conditions for high-density fermentation of S. fibuligera Y1402 through single-factor design, Plackett-Burman design, steepest ascent test, and response surface analysis. The study found that glucose at 360.61 g/L, peptone at 50 g/L, yeast extract at 14.65 g/L, KH2PO4 at 5.49 g/L, MgSO4 at 0.40 g/L, and CuSO4 at 0.01 g/L were the best medium ingredients for S. fibuligera Y1402. Under these conditions, after three days of fermentation, the total colony count reached 1.79 × 108 CFU/mL. The optimal fermentation conditions were determined to be an initial pH of 6.0, an inoculum size of 1.10%, a liquid volume of 12.5 mL/250 mL, a rotation speed of 120 r/min, a fermentation temperature of 21 °C and a fermentation time of 53.50 h. When fermentation was conducted using the optimized medium and conditions, the total colony count achieved a remarkable value of 5.50 × 109 CFU/mL, exhibiting a substantial increase of nearly 31 times the original value in the optimal culture medium. This significant advancement offers valuable insights and a reference for the industrial-scale production of S. fibuligera.

Screening, Identification, and Fermentation Condition Optimization of a High-Yield 3-Methylthiopropanol Yeast and Its Aroma-Producing Characteristics.

A high-yield 3-methylthiopropanol (3-Met) yeast Y1402 was obtained from sesame-flavored Daqu, and it was identified as Saccharomycopsis fibuligera. S. fibuligera Y1402 showed a broad range of growth temperatures and pH, as well as the maximum tolerance to glucose, NaCl, nicotine, and 3-Met at 50% (w/w), 15% (w/v), 1.2 g/L, and 18 g/L, respectively. After optimization using single-factor experiments, a Plackett-Burman design, a steepest ascent test, and a Box-Behnken design, the 3-Met yield reached 4.03 g/L by S. fibuligera Y1402 under the following optimal conditions: glucose concentration of 40 g/L, yeast extract concentration of 0.63 g/L, Tween 80 concentration of 2 g/L, L-methionine concentration of 5 g/L, liquid volume of 25 mL/250 mL, initial pH of 5.3, fermentation temperature of 32 °C, inoculum size of 0.8%, shaking speed of 210 rpm, and fermentation time of 54 h. The fermentation was scaled up to a 3 L fermenter under the optimized conditions, and the yield of 3-Met reached 0.71 g/L. Additionally, an aroma analysis revealed that the flavor substances produced by S. fibuligera Y1402 in sorghum hydrolysate medium was mainly composed of compounds with floral, sweet, creamy, roasted nut, and clove-like aromas. Therefore, S. fibuligera has great potential for application in the brewing of Baijiu and other fermented foods.

Polarization-insensitive antisymmetric multimode waveguide Bragg grating filter based on an SiN-Si dual-layer stack.

A polarization-insensitive multimode antisymmetric waveguide Bragg grating (MASWBG) filter based on an SiN-Si dual-layer stack is demonstrated. Carefully optimized grating corrugations patterned on the sidewall of a silicon waveguide and SiN overlay are used to perturbate TE and TM modes, respectively. Furthermore, the lateral-shift apodization technique is utilized to improve the sidelobe suppression ratio (SLSR). A good overlap between the passbands measured in TE and TM polarization states is obtained. Insertion losses, SLSRs, and 3-dB bandwidths of measured passbands in TE/TM polarizations are 1/1.72 dB, 18.5/19.1 dB, and 5.1/3.5 nm, respectively.

Silicon-based high-power traveling wave photodetector with inductive gain peaking.

We demonstrate Ge/Si high-power and high-speed distributed traveling wave photodetectors (TWPD) by using the inductive gain peaking technique. Input terminals of TW electrodes are open to enhance RF output efficiencies to output loads. Furthermore, optimized on-chip spiral inductors are incorporated at output terminals of TW electrodes to alleviate bandwidth degradations caused by the absences of matching impedances. A comprehensive equivalent circuit model is developed to calculate the frequency response of this scheme. It is used to optimize the design, and then is validated by measurement results. After inducing on-chip inductors, the bandwidths of 4-stage and 8-stage TWPDs are improved from 32 to 44 GHz and 16 to 24 GHz, respectively. Maximum RF output powers of 4-stage and 8-stage TWPDs with on-chip inductors are measured to be 5.7 dBm and 9.4 dBm at 20 GHz, respectively.

High linearity silicon DC Kerr modulator enhanced by slow light for 112 Gbit/s PAM4 over 2 km single mode fiber transmission.

We demonstrate a high efficiency, high linearity and high-speed silicon Mach-Zehnder modulator based on the DC Kerr effect enhanced by slow light. The two modulation arms based on 500-µm-long grating waveguides are embedded with PN and PIN junctions, respectively. A comprehensive comparison between the two modulation arms reveals that insertion loss, bandwidth and modulation linearity are improved significantly after employing the DC Kerr effect. The complementary advantages of the slow light and the DC Kerr effect enable a modulation efficiency of 0.85 V·cm, a linearity of 115 dB·Hz2/3, and a bandwidth of 30 GHz when the group index of slow light is set to 10. Furthermore, 112 Gbit/s PAM4 transmission over 2 km standard single mode fiber (SSMF) with bit error ratio (BER) below the soft decision forward error correction (SD-FEC) threshold is also demonstrated.

Clostridium btbubcensis BJN0001, a potentially new species isolated from the cellar mud of Chinese strong-flavor baijiu, produces ethanol, acetic acid and butyric acid from glucose.

A novel strain, designated BJN0001, was isolated from the cellar mud of Chinese strong-flavor baijiu. The complete genome of strain BJN0001 was 2,688,791 bp and annotated with 2610 genes. Whole-genome similarity metrics such as average nucleotide identity (ANI) of BJN0001 with reference genomes reveals clear species boundaries of < 95% ANI value for species. The DNA-DNA hybridization (DDH) values of BJN0001 with the type species were all lower than 70% DDH value for species. Based on these results, the strain BJN0001 was considered a potentially new species of the genus Clostridium. Meanwhile, the fermentation characteristics indicated that the strain had the capability to convert glucose to ethanol, acetic acid and butyric acid, which could provide basic data for revealing its function in baijiu fermentation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03271-7.

Ultra-compact silicon mode (de)multiplexer based on directional couplers with subwavelength sidewall corrugations.

Asymmetrical directional couplers aided with subwavelength sidewall corrugations are used to realize ultra-compact silicon mode (de)multiplexers at C-band. Three mode (de)multiplexers with ultra-short coupling lengths of 5.6/6.5/7.7 µm are designed to enable low-loss mode conversions between TE0 and TE1/2/3 modes. They are then cascaded to build a four-channel mode-division-multiplexing (MDM) link. The four mode channels present minimal on-chip insertion losses of 0.2/0.7/0.7/0.9 dB at their peak wavelengths. Measured cross talk levels of the four mode channels are better than -18.0/-19.1/-16.0/-18.2 dB within the wavelength range from 1530 nm to 1580 nm.

Biochemical characterization of a novel feruloyl esterase from Burkholderia pyrrocinia B1213 and its application for hydrolyzing wheat bran.

In this study, a novel feruloyl esterase (BpFae) from Burkholderia pyrrocinia B1213 was purified, biochemically characterized, and applied in releasing ferulic acid from wheat bran. The molecular mass of BpFae was approximately 60 kDa by SDS-PAGE, and the enzyme was a homomultimer in solution. BpFae displayed maximum activity at pH 4.5-5.0 and was stable at pH 3.0-7.0. The optimal temperature for BpFae was 50 °C. BpFae activity was not affected by most metal ions tested and was significantly increased by Tween-20 and Triton-100. Purified BpFae exhibited a preference for methyl ferulate (41.78 U mg-1) over methyl p-coumarate (38.51 U mg-1) and methyl caffeate (35.36 U mg-1) and had the lowest activity on methyl sinapate (1.79 U mg-1). Under the optimum conditions, the K m and V max for methyl ferulate were 0.53 mM and 86.74 U mg-1, respectively. Residues Ser209, His492, and Glu245 in the catalytic pocket of BpFae could form hydrogen bonds with the substrate and were crucial for catalytic activity and substrate specificity. When G11 xylanase XynA and BpFae were used separately for hydrolyzing de-starched wheat bran (DSWB), the ferulic acid released was undetectable and 1.78%, respectively, whereas it was increased to 59.26% using the mixture of the two enzymes. Thus, BpFae is considered an attractive candidate for the production of ferulic acid from agricultural by-products. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03066-2.

Correlation between microbial communities and flavor compounds during the fifth and sixth rounds of sauce-flavor baijiu fermentation.

Sauce-flavor baijiu is a representative of Chinese traditional fermented baijiu using grains as the raw materials through the co-fermentation of microorganisms. The whole manufacturing process includes 7 times of distillation and generates 7 kinds of base baijius. The final product is a mixture of the 7 kinds of base baijius. Thus the base baijius greatly affect the quality of the final product. The quality of the base baijiu obtained by the sixth distillation is obviously poorer than that of the fifth one. However, the reason is still unclear and limits the quality control of baijiu fermentation. In this study, the flavor substances and microbiota in the up, middle and bottom layers of fermented grains in the fifth and sixth rounds were compared. Some flavor esters showed obviously decreased concentrations in the sixth round, including ethyl benzoneacetic acid, ethyl hexanoic acid, ethyl dodecanoic acid, diethyl butanedioic acid, and ethyl 2-hydroxyl-propanoic acid. Meanwhile, an off-flavor p-cresol was detected in the sixth round. Correlation analysis of flavor chemicals and microbiota indicated that fungi in the fifth round played an important role for ester synthesis. Some bacterial and fungal species were both positively correlated with p-cresol synthesis, and the related p-cresol metabolic pathways were proposed for the first time. These results revealed flavor divergences of fermented grains between the fifth and sixth rounds, and will ultimately help to improve baijiu quality.

Polarization-independent fiber-chip grating couplers optimized by the adaptive genetic algorithm.

One-dimensional polarization-independent grating couplers are demonstrated with the aid of the adaptive genetic algorithm optimization. By adjusting the relative weight between the coupling efficiency and the bandwidth of the polarization-dependent loss (PDL), we control the evolution direction and customize the final performance of the device. Two specific designs are generated by giving more weight to the coupling efficiency and the PDL bandwidth, respectively. Coupling efficiencies of the first design are measured to be -7.6dB and -7.9dB at 1550 nm for TE and TM polarizations, respectively, while its 1.0 dB PDL bandwidth is 25.0 nm. In contrast, the second design presents higher coupling efficiencies of -7.6dB and -7.2dB at 1550 nm for TE and TM polarizations, respectively. However, its 1.0 dB PDL bandwidth is 22.0 nm.

Application of Wickerhamomyces anomalus in Simulated Solid-State Fermentation for Baijiu Production: Changes of Microbial Community Structure and Flavor Metabolism.

Wickerhamomyces anomalus is conducive to the synthesis of ester compounds in brewing the Chinese liquor Baijiu; esters are crucial for the quality of Baijiu. In this study, simulated solid-state fermentation for Baijiu production was used to explore whether artificial addition of W. anomalus could improve the flavor substance in Baijiu, and the underlying mechanisms. Two experimental groups were studied, in which W. anomalus Y3604 (Group A) and YF1503 (Group B) were added, respectively; in the control group (Group C), no W. anomalus was added. Adding strain Y3604 increased the content of esters in fermentation samples, especially ethyl acetate and ethyl caproate, and reduced the content of higher alcohols. Adding strain YF1503 had little effect on the ester content but decreased the content of higher alcohols. The diversity and abundance of prokaryotic genera in Group A and B samples were similar, but there were some differences compared with Group C. The correlations of genera in Group A or B samples were simple compared with group C. Although the predominant eukaryotic genera in the three groups were consistent, the abundance of each gene varied among groups. Based on our findings, bioaugmentation of Baijiu fermentation with W. anomalus will change the ethyl acetate content and cause changes in the levels of other flavor substances. We suggest that the changes in flavor substances caused by the addition of W. anomalus are mainly due to changes in the microbial community structure that result from the addition of W. anomalus.

Optimized soluble expression of a novel endoglucanase from Burkholderia pyrrocinia in Escherichia coli.

Burkholderia pyrrocinia B1213, a novel microbe isolated from a Baijiu-producing environment, displayed strong cellulolytic activity on agar plates with glucan as the carbon source and had an activity of 674.5 U/mL after culturing with barley. Genome annotation of B. pyrrocinia identificated a single endoglucanase (EG)-encoding gene, designated as BpEG01790. The endoglucanase BpEG01790 shows 98.28% sequence similarity with an endo-ß-1,4-glucanase (EC 3.2.1.4) from Burkholderia stabilis belonging to glycoside hydrolase family 8 (GH8). The gene BpEG01790 has an open reading frame of 1218 bp encoding a 406 amino acid (AA) residue protein (43.0 kDa) with a 40-AA signal peptide. BpEG01790 was successfully cloned into pET28a( +) with and without the signal peptide; however, attempts to overexpress this protein in Escherichia coli BL21(DE3) cells using this expression system failed. BpEG01790 was also cloned into the pCold TF vector. Active BpEG01790 was successfully overexpressed with or without the signal peptide using the pCold TF vector expression system and E. coli BL21 (DE3) cells. Overexpression of recombinant BpEG01790 without the signal peptide was higher compared with the construct that included the signal peptide. Optimization of culture conditions improved the enzyme activity by 12.5-fold. This is the first report describing the heterologous soluble overexpression of an EG belonging to GH8 from B. pyrrocinia using TF as a molecular chaperone.

High linearity silicon modulator capable of actively compensating input distortion.

In this Letter, we demonstrate an ultra-high linearity silicon carrier-depletion-based modulator by integrating a dual-parallel Mach-Zehnder modulator (DP-MZM) with a 1×2 thermo-optical switch. The operation principle is to manipulate power distributions of RF and optical signals among the two sub-MZMs, so their third-order nonlinearities can cancel each other. Spurious-free dynamic ranges (SFDRs) for the third-order intermodulation distortion (IMD3) are measured to be 123/120dB⋅Hz6/7 at 1/10 GHz, which represents a record-high linearity achieved with silicon-based modulators. As a contrast, SFDRs of a reference single MZM are 113/108dB⋅Hz4/5 at the same frequencies. Furthermore, we first demonstrate that this device is able to actively compensate nonlinear distortions of RF driving signals in the optical domain. Carrier-to-distortion ratios (CDRs) of deliberately distorted two-tone signals are improved from 40/50 dB to 45/72 dB after the modulation.

Screening of yeasts isolated from Baijiu environments for 2-phenylethanol production and optimization of production conditions.

2-Phenylethanol (2-PE) with a pleasant rose-like odor is a valuable aroma compound used in many fields. 2-PE production by yeast is considered a promising alternative to chemical synthesis and extraction from natural materials. In this report, the strain YF1702 produced a significantly higher level of 2-PE when compared with other strains isolated from Baijiu-producing environments. According to morphological properties, physiological and biochemical characteristics, and 26S rDNA sequence analysis, strain YF1702 was identified as Pichia kudriavzevii. The optimal fermentation conditions of YF1702 for producing 2-PE were obtained by single-factor experiments, Plackett-Burman design, steepest ascent design, and response surface methodology. The optimal inoculation conditions for strain YF1702 were 50 g/L glucose, 6.0 g/L yeast extract, 10.7 g/L L-Phe, and 32 g/L Tween-60. The optimal fermentation conditions were pH 2.3, 26 °C, 210 rpm shaking, an inoculum size of 0.4% (v/v), and a loading volume of 25.5 mL/250 mL for 56 h. Under these optimal conditions 2-PE production by YF1702 was 5.09 g/L. This strain has the potential to increase the content of 2-PE in Baijiu production and enhance the aroma characteristics of Baijiu.

Hitless and gridless reconfigurable optical add drop (de)multiplexer based on looped waveguide sidewall Bragg gratings on silicon.

Reconfigurable optical add-drop filters in future intelligent and software controllable wavelength division multiplexing networks should support hitless wavelength switching and gridless bandwidth tuning. The hitless switching implies that the central wavelength of one channel can be shifted without disturbing data transmissions of other channels, while the gridless tuning means that the filter bandwidth can be adjusted continuously. Despite a lot of efforts, very few integrated optical filters simultaneously support the hitless switching of central wavelength and the gridless tuning of bandwidth. In this work, we demonstrate a hitless add-drop filter with gridless bandwidth tunability on the silicon-on-insulator (SOI) platform. The filter comprises the two identical multimode anti-symmetric waveguide Bragg gratings (MASWBG) which are connected to a loop. The phase apodization technique is utilized to weaken the intrinsic sidelobe interference of grating-based devices. By sequentially manipulating central wavelengths of the two MASWBGs with the thermo-optical effect, we can reconfigure the spectral response of the filter gridlessly and hitlessly. Specifically, the central wavelength of the device is shifted by 14.5 nm, while its 3 dB bandwidth is tuned from 0.2 nm to 2.4 nm. The dropping loss and the sidelobe suppression ratio (SLSR) are dependent on the bandwidth selected. Measured variation ranges of dropping loss and SLSR are from -1.2 dB to -2.5 dB and from 12.8 dB to 21.4 dB, respectively. The hitless wavelength switching is verified by a data transmission measurement at a bit rate of 25 Gbps.

Optimization of fermentation conditions for the production of recombinant feruloyl esterase from Burkholderia pyrrocinia B1213.

Statistical experimental designs were used to optimize conditions for recombinant Burkholderia pyrrocinia feruloyl esterase (BpFae) production in bacteria under lactose induction. After optimization by single factor design, Plackett-Burman design, steepest ascent design and the response surface method, the optimal conditions for BpFae production were: 6 g/L lactose, pH 5.5, pre-induced period 5 h, 23 °C, shaker rotational speed of 240 rpm, medium volume of 50 mL/250 mL, inoculum size 0.2% (v/v), and a post-induced period of 32 h in a Luria-Bertani culture. The produced BpFae activity was 7.43 U/mL, which is 2.92 times higher than that obtained under optimal conditions using IPTG as the inducer. BpFae activity was 4.82 U/mL in a 5 L fermenter under the abovementioned optimal conditions. BpFae produced a small amount of ethyl acetate but had no effect on the synthesis of other important esters in Baijiu. The results underpin further investigations into BpFae characterization and potential applications.

Effects of aging on the quality of roasted sesame-like flavor Daqu.

BACKGROUND: Daqu, the saccharification, fermentation, and aroma-producing agents for Baijiu brewing, is prepared using a complex process. Aging is important for improving the quality of Daqu, but its impact has rarely been studied. This study investigated changes in the physicochemical properties, flavor compounds, and microbial communities during aging of Daqu with a roasted sesame-like flavor. RESULTS: The physicochemical properties changed continuously during aging to provide a high esterifying activity. Aging removed unpleasant flavor compounds and helped to stabilize the flavor compounds in mature Daqu. A high-throughput sequencing approach was used to analyze the changing composition of the microbial communities during aging. Aging helped to modify the microbial population to produce better Baijiu by eliminating low-abundance microbial communities and optimizing the proportion of predominant microbial communities. Nine genera of prokaryotic microbes formed the core microbiota in Daqu after aging. Regarding eukaryotic microbes, Zygomycota, the predominant community, increased in the first 2 months, then decreased in the third month of aging, while Ascomycota, the subdominant community, showed the opposite behavior. Absidia, Trichocomaceae_norank and Rhizopus were the predominant genera in the mature Daqu. CONCLUSIONS: Significant correlations between microbiota and physicochemical properties or flavor compounds were observed, indicating that optimizing microbial communities is essential for aging Daqu. This study provides detailed information on aging during Daqu preparation.

High-performance silicon-graphene hybrid plasmonic waveguide photodetectors beyond 1.55 µm.

Graphene has attracted much attention for the realization of high-speed photodetection for silicon photonics over a wide wavelength range. However, the reported fast graphene photodetectors mainly operate in the 1.55 µm wavelength band. In this work, we propose and realize high-performance waveguide photodetectors based on bolometric/photoconductive effects by introducing an ultrathin wide silicon-graphene hybrid plasmonic waveguide, which enables efficient light absorption in graphene at 1.55 µm and beyond. When operating at 2 µm, the present photodetector has a responsivity of ~70 mA/W and a setup-limited 3 dB bandwidth of >20 GHz. When operating at 1.55 µm, the present photodetector also works very well with a broad 3 dB bandwidth of >40 GHz (setup-limited) and a high responsivity of ~0.4 A/W even with a low bias voltage of -0.3 V. This work paves the way for achieving high-responsivity and high-speed silicon-graphene waveguide photodetection in the near/mid-infrared ranges, which has applications in optical communications, nonlinear photonics, and on-chip sensing.

Effects of fortification of Daqu with various yeasts on microbial community structure and flavor metabolism.

Fortification of Daqu with isolated functional strains can influence the metabolic activity of the microbial community, and thus alter the flavors of the Baijiu produced with Daqu as a fermentation starter. Here, we analyzed the microbial community dynamics of, and volatile compound production by, Daqu fortified respectively with three high-yield ethyl caproate-producing yeasts (Saccharomyces cerevisiae Y7#09, Hyphopichia burtonii F12507 and Clavispora lusitaniae YX3307), or with a mixture of these three strains, during the fermentation of Baijiu. The microbial community was investigated using Illumina HiSeq technology. Three bacterial genera (Bacillus, Lactobacillus and Enterobacter) and four fungal genera (Pichia, Clavispora, Saccharomyces and Saccharomycopsis) were dominant in the microbial communities. The volatile compounds were examined by gas chromatography-mass spectrometry. Forty-one flavor compounds were detected in all samples, including seven alcohols, 26 esters and four aldehydes. In particular, an increase in ethyl caproate content was associated with Daqu fortified with S. cerevisiae Y7#09, C. lusitaniae YX3307, or the mixed inoculum. The ester content of these fortified Daqu was higher in the later stage of the fermentation than that in unfortified Daqu, or in Daqu fortified with H. burtonii F12507. Our results show that fortification of Daqu with aroma-producing yeast strains influenced the microbial community composition in the Daqu and affected its metabolic activity. Overall, this study reveals the features of fortified Daqu microbial communities in different phases and improves understanding of the relationships between aroma-producing yeast and the metabolic activity of microbial communities in Baijiu production.