**4. Materials and Methods**

#### *4.1. General*

All chemicals were purchased from Sigma–Aldrich (Schnelldorf, Germany) and used without further purification unless specified otherwise. The culture media components were obtained from BD (Becton, Dickinson and Company, Bremen, Germany).

1H and 13C NMR spectra were recorded using a Bruker Advance 400 (Karlsruhe, Germany) (400 MHz and 100 MHz, respectively) instrument and internally referenced to residual solvent signals. Data for 1H NMR are reported as chemical shift (d ppm), multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet), integration, coupling constant (Hz) and assignment. Data for 13C NMR are reported in terms of chemical shift. Optical rotations were obtained at 20 ◦C using a PerkinElmer 241 polarimeter (Shanghai, China) (sodium D line). Column chromatography was performed with a silica gel (0.060–0.200 mm, pore diameter ca. 6 nm) and mixtures of petroleum ether (PE) and ethyl acetate (EtOAc) as solvents. Thin-layer chromatography (TLC) was performed on 0.20 mm silica gel 60-F plates. Organic solutions were concentrated under reduced pressure with a rotary evaporator.

#### *4.2. Chemical Synthesis of the Standard Racemic β-Phenylalcohols* **2a***–***2f***,* **2m** *and* **2n**

Ten mmol of NaBH4 was added to a cooled (0 ◦C) solution of 2.5 mmol of each specific substrate (**1a**–**1f**, **1m** and **1n**) in 50 mL of methanol. After stirring for 10 min, the mixture was warmed to room temperature and stirred for another 3–4 h to complete the reduction. After quenching with 2 M HCl to pH 7.0, the mixture was extracted with EtOAc (50 mL × 3). The organic phases were washed with brine, dried over Na2SO4, filtered and concentrated in vacuum. The residue was purified by flash chromatography on silica gel (eluent: EtOAc/PE 1:20) to give the racemic alcohol **2a**–**2f**, **2m** and **2n** (see Supplementary Materials for NMR spectroscopic data).

#### *4.3. Microorganisms*

The marine fungi strains *Penicillium citrinum* GIM 3.458, *Penicillium citrinum* GIM 3.251, *Penicillium citrinum* GIM 3.100, *Aspergillus sclerotiorum* AS 3.2578, *Aspergillus sydowii* AS 3.7839, *Aspergillus sydowii* AS 3.6412, *Geotrichum candidum* GIM 2.361, *Geotrichum candidum* GIM 2.616, *Rhodotorula rubra* GIM 2.31, *Rhodotorula mucilaginosa* GIM 2.157, *Geotrichum candidum* AS 2.1183, *Geotrichum candidum* AS 2.498 and *Rhodotorula rubra* AS 2.2241 were isolated from a wide collection of isolates from marine sediments from Guangdong Province, China. All strains used in this study were deposited and commercially available at Guangdong Culture Collection Center or the China General Microbiological Culture Collection Center.

The marine fungi were maintained on agar plates at 4 ◦C and subcultured at regular intervals. The medium (RM1) used for cultivation contained glucose (15 g/L), peptone (5 g/L), yeast extract (grease, 5 g/L), disodium hydrogen phosphate (0.5 g/L), sodium dihydrogen phosphate (0.5 g/L), magnesium sulphate (0.5 g/L) and sodium chloride (10 g/L) and a final pH 7.0; this medium was sterilized at 115 ◦C in an autoclave for 25 min. A loop of a single colony was cut from the agar stock cultures and used to inoculate a given medium in an appropriate Erlenmeyer flask. This culture was shaken reciprocally (220 ppm) at 28 ◦C for given times.

#### *4.4. Transformation with Growing Cells*

Growing cells of *R. rubra* AS 2.2241 were used for the transformation of 1-(3-bromophenyl)ethanone (**1b**). 1-(3-bromophenyl)ethanone (**1b**) was added at the time of inoculation of the RM1 medium as described above by *R. rubra* AS 2.2241; in another set of experiments, 1-(3-bromophenyl)ethanone (**1b**) was added to the growing cells at different phases of growth, and the transformation continued.

#### 4.4.1. **1b** Was Added at the Time of Inoculation

Different concentrations (5–15 mM) of 1-(3-bromophenyl)ethanone (**1b**) were added aseptically to 500-mL Erlenmeyer flasks containing 250 mL of sterile RM1 medium (pH 7.0). Three control flasks were prepared. The first control was prepared with 10 mM 1-(3-bromophenyl)ethanone in 250 mL of RM1 medium, and the flask was not inoculated with *R. rubra* AS 2.2241. This was used as a positive control for 1-(3-bromophenyl)ethanone (**1b**). The second control contained only the RM1 medium without 1-(3-bromophenyl)ethanone (**1b**), and the flask was inoculated with *R. rubra* AS 2.2241. This was done to check the growth of *R. rubra* AS 2.2241 in the absence of 1-(3-bromophenyl)ethanone (**1b**). The third control was prepared with 5 g of (dry weight) dead cells of *R. rubra* AS 2.2241 in RM1 medium with 10 mM 1-(3-bromophenyl)ethanone (**1b**). This was used to determine the active enzymes by the cells. This flask was not inoculated. All flasks were kept in a temperature-controlled (28 ◦C) orbital shaker at 220 rpm shaking speed. The samples were analysed after 24 h. The cells were removed by centrifugation at 4000 rpm and at 4 ◦C for 20 min. The collected cells were used for cell mass analysis. The supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

#### 4.4.2. **1b** Were Added at Different Phases of Growth

Several 500-mL Erlenmeyer flasks containing 250 mL of RM1 medium (pH 7.0) were inoculate with *R. rubra* AS 2.2241 and cultured at 28 ◦C on an orbital shaker (220 rpm). Next, 10 mM 1-(3-bromophenyl)ethanone (**1b**) was added to the growing cells after 24, 48, 72, 96 h of inoculation in separated flasks. For the transformation reaction, the flasks were again incubated under the same conditions. The samples were withdrawn at regular intervals and centrifuged, and the supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

#### *4.5. General Methods for Growing Cells Biotransformation*

All substrates **1a**–**1n** were added at the time of inoculation, and the reaction setup was the same as "**1b** was added at the time of inoculation". After a 72-h inoculation, the reaction was stopped by centrifugation at 4000 rpm and at 4 ◦C for 20 min to remove the cells. The supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

#### *4.6. Transformation with Resting Cells*

*R. rubra* AS 2.2241 was cultivated in RM1 medium (pH 7.0) at 28 ◦C with 220 rpm speed. Cells in the culture age of 24 h, 48 h, 72 h and 96 h were harvested by centrifugation and washed twice with 100 mM Na2HPO4-KH2PO4 buffer (pH 7.0). Approximately 3 g of resting cells of 24-h-grown age was suspended in 10 mL of Na2HPO4-KH2PO4 buffer with the required pH (pH 6.0, 7.0 and 8.0) containing 0.5 g of glucose and 10 mM 1-(3-bromophenyl)ethanone (**1b**). The reaction mixtures were shaken (220 rpm) at the given temperatures (20 ◦C, 25 ◦C and 35 ◦C) for 24 h and stopped by centrifugation at 4000 rpm and at 4 ◦C for 20 min to remove the cells. The supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

Resting cells of 48 h-, 72 h- and 96-h-grown ages were also suspended in 10 mL Na2HPO4-KH2PO4 buffer (pH 7.0, 100 mM) containing 0.5 g glucose and 10 mM of 1-(3-bromophenyl)ethanone (**1b**). The reaction mixtures were shaken (220 rpm) at 25 ◦C for 24 h and stopped by centrifugation at 4000 rpm and at 4 ◦C for 20 min to remove cells. The supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

#### *4.7. Recyclability*

Reactions were carried out with 10 mM substrate **1b** in 10 mL of Na2HPO4-KH2PO4 buffer (100 mM, pH 7.0) containing 0.5 g of glucose and 3 g of resting cells of 48-h-grown age, shaken at 25 ◦C for 24 h. At the end of the reaction, the cells were centrifuged at 4000 rpm for 20 min to be separated from the reaction mixture, then washed by Na2HPO4-KH2PO4 buffer (100 mM, pH 7.0) and resuspended in 10 mL of the same buffer containing the same substrates and glucose. The reaction mixture (2 mL of supernatant separated from cells) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.

### *4.8. General Methods for Resting Cell Biotransformation*

Reactions were performed in 50-mL screw-capped glass vials to prevent evaporation of the substrate/product. Shaking was performed in a heated ground-top shaker at 25 ◦C with 220 rpm. Approximately 3 g of resting cells of 48-h-grown age (wet cells) were resuspended in 10 mL of Na2HPO4-KH2PO4 buffer (100 mM, pH 7.0) containing 0.5 g of glucose and 10 mM **1a**–**1n**. After 24 h, the reaction was stopped by centrifugation at 4000 rpm and at 4 ◦C for 20 min to remove cells. The supernatant (2 mL) was saturated with NaCl followed by extraction with 2 × 2 mL of HPLC eluent (*n*-hexane/*i*-PrOH = 95/5, *v/v*) by shaking for 5 min. The combined organic layer was dried over Na2SO4 and measured by HPLC for yield and *ee*.
